Data_Sheet_1_Performance-guaranteed distributed control for multiple plant protection UAVs with collision avoidance and a directed topology.docx

The urgent requirement for improving the efficiency of agricultural plant protection operations has spurred considerable interest in multiple plant protection UAV systems. In this study, a performance-guaranteed distributed control scheme is developed in order to address the control of multiple plant protection UAV systems with collision avoidance and a directed topology. First, a novel concept called predetermined time performance function (PTPF) is proposed, such that the tracking error can converge to an arbitrary small preassigned region in finite time. Second, combined with the two-order filter for each UAV, the information estimation from the leader is generated. The distributed protocol avoids the use of an asymmetric Laplace matrix of a directed graph and solves the difficulty of control design. Furthermore, by introducing with a collision prediction mechanism, a repulsive force field is constructed between the dynamic obstacle and the UAV, in order to avoid the collision. Finally, it is rigorously proved that the consensus of the multiple plant protection UAV system can be achieved while guaranteeing the predetermined time performance. A numerical simulation is carried out to verify the effectiveness of the presented method, such that the multiple UAVs system can fulfill time-constrained plant protection tasks.</p

AIB1 regulates the morphologies of breast cancer cells.

<p>(A) Light microscopic images showing the morphology of different breast cancer cells. (B) Expression levels of AIB1, ERα and E-cadherin in different breast cancer cells as detected by western blot analysis. Cell extracts were prepared from the different cell lines and probed with specific antibody against AIB1, ERα and E-cadherin or β-actin. (C) Light microscopic images showing EMT morphological changes induced in T47D cells after treatment with EGF (50 ng/ml) or E2 (20 nM) for 24 h. (D) Western blot analysis showing the levels of AIB1, ERα, E-cadherin and β-actin expressions in T47D cells after treatment with EGF or E2. (E) Coimmunoprecipitation showing AIB1 and ERα complex increased after treatment with EGF or E2. T47D cells treated without or with EGF (50 ng/ml) or E2 (20 nM) for 24 h were subjected to immunoprecipitation with anti-AIB1 or control IgG antibodies, followed by western blot analysis with anti-ERα and anti-AIB1 antibodies. (F) Light microscopic images showing T47D cells without or with AIB1 knockdown (shAIB1#1 and shAIB1#2), ERα knockdown (shERα) or both AIB1 and ERα knockdown. Cells were treated with the corresponding iRNA and then plated out in 50-mm dishes and incubated for 3 days before observing. (G) The cells from (F) were counted and plotted as percentage of clustered or scattered cells relative to total number of cells (400–500). (H) Western blot analysis showing the levels of AIB1, ERα, E-cadherin and β-actin proteins in T47D cells from (F).</p

AIB1 regulates ERα-mediated SNAI1 expression.

<p>(A) Schematic illustration of ERα-binding elements in SNAI1 promoter. Fragments A to C were chosen for PCR amplification in ChIP assays. Three truncated versions of the SNAI1 promoter were made, and the length of each is shown in the illustration. (B) ChIP assays showing AIB1- and ERα-SNAI1 promoter interaction in T47D cells. Cross-linked chromatin was extracted from T47D cells and subjected to immunoprecipitation with anti-AIB1, anti-ERα or control IgG, and the resulting precipitated DNA was used as template for PCR-ampαlification of SNAI1 promoter using specific primer covering region A, B or C of the promoter region. (C) Reporter gene assays of different truncated versions of SNAI1 promoter in the presence of AIB1 or ERαoverexpression. Each of the truncated SNAI1 promoters was fused to luciferase gene in pGL3 and the resulting construct was introduced into T47D cells along with AIB1 or ERα construct or both. The levels of luciferase activity in these cells were determined 48 h after transfection. Luciferase activity was normalized to β-galactosidase activity, used to evaluate transfection efficiency. Each experiment was performed in triplicates and repeated at least of three times. Data are the means ± SDs. Statistically significant differences (<i>P</i><0.05) in paired Student’s t-test are marked with an asterisk.</p

AIB1 promotes cell motility and invasion in T47D cells.

<p>(A) Scratch wound-healing assay showing the effect of AIB1 on cell motility in T47D cells. Top panel: Images of T47D cells that were transfected with empty vector (pcDNA) or AIB1, and cells without (NC) or with AIB1 knockdown (shAIB1#1) before and after wound-healing assay. The cell layers were carefully wounded using a sterile 200-µl tip and then cultured for 24 h before evaluation. Bottom panel: Western blot analysis showing the levels of AIB1, ERα and β-actin expressions in pcDNA- or AIB1-transfected T47D cells, and of T47D cells without or with AIB1 knockdown. (B) Transwell migration and invasion assays showing the effect of AIB1 on cell motility and invasion ability in T47D cells. Images showing the migration and invasion of T47D cells that were transfected with empty vector (pcDNA) or AIB1, and of T47D cells without (NC) or with AIB1 knockdown (shAIB1#1). For NC group, the cells were transfected with a negative control scrambled shRNA synthesis DNA cloned into siRNA expression vector pRNAT carries GFP marker. Cell migration and invasion assays were performed in 24-well chambers without and with Matrigel, respectively. Cells (1000 per well) were transfected with GFP-AIB1 or just GFP (pEGFPC) and then transferred to the upper chamber. After 48 h of incubation, the numbers of migrating and invasive cells on the lower surface of the filter were counted under a fluorescent microscope. The bar graphs on the right of the images show the number of migrating and invading cells for each category of cells. (C) MTT assays showing the effect of AIB1 on cell proliferation in T47D cells. Cells were transfected as in (A) and then subjected to MTT assay within 24 hours.</p

AIB1 regulates E-cadherin expression through E-box-dependent transcription.

<p>(A) Western blot analysis of E-cadherin and N-cadherin levels in T47D and MCF-7 cells that overexpressed AIB1 or had AIB1 knockdown. Cells were transfected with either empty vector (pcDNA) or AIB1, or with the negative control scrambled shRNA (NC) or shAIB1#1 (in the case of AIB1 knockdown). Western blot analysis was carried out using cell extract and antibody against AIB1, E-cadherin, N-cadherin or β-actin. (B) Immunofluorescence study showing the regulation of AIB1 on the expression of E-cadherin and N-cadherin. Cells overexpressed AIB1 and empty vector or AIB1 knockdown and negative control scrambled shRNA (NC) were cultured in the chamber slide and fixed, immunostained with anti-E-cadherin and anti-N-cadherin antibodies followed by the secondary antibody, Texas-red Fluor 589 anti-Rabbit. Nuclear protein was stained with DAPI. (C) Fluorescence microscopic images showing the morphology of T47D and MCF-7 cells that overexpressed AIB1 and the empty vector pEGFPC1. (D) RT-PCR analysis showing the regulation of AIB1 on the transcription of E-cadherin. The mRNA level of E-cadherin was expressed relative to GAPDH transcript level. (E) AIB1-regulated E-cadherin luciferase reporter activity. T47D cells were co-transfected with the E-cadherin-Luc reporter vector (pGL3-Ecad) and AIB1 or empty vector. Cells were harvested 48 h after transfection and subjected to luciferase activity assay. (F) Dependence of AIB1-suppressed E-cadherin transcription on E-box as shown by luciferase reporter activity. T47D cells were cotransfected with empty vector (pcDNA) or vector plus AIB1 insert (pcDNA-AIB1) together with the E-box wild-type (pGL3-Ecad) or E-box mutant (pGL3-Ecad-mut) E-cadherin-Luc reporter. Cells were harvested 48 h after transfection and subjected to luciferase activity. Relative luciferase activity was normalized to β-galactosidase activity, used as control to monitor the transfection efficiency. Each experiment was performed in triplicates and repeated at least three times. Data are the means ± SDs. Statistically significant differences (<i>P</i><0.05) in paired Student’s t-test are marked with an asterisk.</p

AIB1 induces SNAI1 expression.

<p>(A) RT-PCR analysis showing the regulation of the expression of EMT-inducing transcription factor by AIB1. The mRNA levels of three EMT-inducing transcription factors, SNAI1, SNAI2 and ZEB1, in T47D cells without (pcDNA) or with AIB1 overexpression (AIB1) or without (NC) and with AIB1 knockdown (shAIB1#1) were measured by RT-PCR. The mRNA levels of SNAI1, SNAI2 and ZEB1 are expressed relative to GAPDH transcripts. (B) AIB1-regulated SNAI1 luciferase reporter activity. T47D cells were cotransfected with SNAI1-Luc and different amounts (0.1–0.5 ng) of pcDNA-AIB1 construct. Luciferase activity level in cells transfected with the empty vector pcDNA was set to 1. (C) Regulation of the activity of SNAI1 promoter by different transcription factors without and with AIB1 overexpression. T47D cells were transfected with SNAI1-Luc reporter construct and the transcription factor Sp1, ERα, E2F or Jun-D construct with or without AIB1 construct. (D) Regulation of the activity of SNAI1 promoter by nuclear receptors without and with AIB1 overexpression. T47D and HEK293T cells were transfected with SNAI1-Luc reporter vector with ERα or AR construct with and without AIB1 construct. (E) Effect of hormone and nuclear receptor inhibitor on AIB1-regulated <i>SNAI1</i> activity. T47D cells were transfected with SNAI1-Luc reporter construct with or without AIB1 knockdown, and cells were then treated with 20 nM E2 or 10 nM ICI (inhibitor of estrogen receptor) or without treatment (control) for 12 h. The levels of luciferase were normalized to β-galactosidase activity, used to evaluate transfection efficiency. Each experiment was performed in triplicates and repeated at least three times. Data are the means ± SDs. Statistically significant differences (P<0.05) in paired Student’s t-test are marked with an asterisk.</p

Correlation between AIB1/SNAI1 expressions and E-cadherin expression in human breast tissue samples.

<p>(A) Representative results of immunohistochemistry of AIB1, SNAI1 and E-cadherin in serial sections of primary tumor and invasive tumor tissues (n = 31) respectively. Each sample was incubated with antibody against ERα, AIB1, SNAI1 or E-cadherin. Positive staining and negative staining are indicated by brown and blue staining, respectively (×400 Magnification). (B) Association between AIB1/SNAI1 expressions and E-cadherin proteins in ERα+ breast cancer tissue samples. Fisher exact test, <i>P</i><0.0001.</p

SNAI1 mediates the role of AIB1 in breast cancer cell EMT.

<p>(A) Western blot analysis showing the reversion of repressed E-cadherin expression in SNAI1-knockdown cells by overexpression of AIB1. T47D cells without or with SNAI1 knockdown were transfected with AIB1 and the levels of SNAI1, E-cadherin and β-actin in the cells were analyzed by western blot using the corresponding antibodies. For comparison, the levels of these proteins in T47D cells without SNAI1 knockdown and AIB1 overexpression were also analyzed. (B) RT-PCR analysis showing the level of SNAI1 or E-cadherin transcript in the different groups of cells in A. The mRNA levels of SNAI1 and E-cadherin are expressed relative to GAPDH transcripts. (C) Effect of SNAI1 knockdown on cell motility in T47D cells. The cell motility of T47D cells without or with SNAI1 knockdown that over expressed AIB1 were evaluated by scratch wound healing assay. The motility of cells without SNAI1 knockdown and AIB1 overexpression was also evaluated for comparison purpose. (D) Effect of SNAI1 knockdown on cell migration and invasion abilities of T47D cells. T47D cells without or with SNAI1 knockdown that over expressed AIB1 were subjected to transwell migration and invasion assays. Cells that migrated through the uncoated filter or invaded the Matrigel-coated filters of the chamber were detected by fluorescence imaging. The graph shows the actual number of migrated and invaded cells for each group.</p