PRL1 and CDC5 synergistically regulate miRNA accumulation.

<p>(A) Morphological phenotypes of Col, <i>cdc5-1</i>, <i>prl1-2</i> and <i>cdc5-1 prl1-2</i>. (B) The abundance of miRNAs is lower in <i>cdc5-1 prl1-2</i> than that in <i>cdc5-1</i> or <i>prl1-2</i>. Small RNAs were detected by Northern Blot. To determine the amount of miRNAs, radioactive signals of miRNAs were normalized to U6 RNA. The number represents the relative abundance compared to Col (set as 1) quantified by three repeats (P<0.05). (C) The abundance of pri-miRNAs is reduced in <i>cdc5-1 prl1-2</i>. The levels of pri-miRNAs in various mutants were determined by qRT-PCR, normalized to <i>UBQUITIN5</i> (<i>UBQ5</i>) and compared with those of Col (set as 1). Standard deviation of three technical replications was shown as error bars. **: P<0.01. (D) miR162b production from <i>pre-miR162b</i> in Col, <i>cdc5-1 prl1-2</i>, <i>cdc5-1</i> and <i>prl1-2</i>. The reaction was stopped at 120 min. The radioactive signals of miR162b were normalized to input. The number represents the relative production in various genotypes compared to Col (set as 1) quantified by three repeats (P<0.05).</p

PRL1 associates with the Pol II and DCL1 complexes.

<p>(A) and (B) Co-immunoprecipitation (Co-IP) between PRL1 and Pol II. Protein extracts from transgenic plants containing PRL1-YFP were incubated with Anti-YFP or anti-RPB2 antibodies to precipitate PRL1-YFP or Pol II. PRL1-YFP and RBP2 were detected with western blot and labeled on the left side of the picture. Ten percent of input proteins were used for IP and one percent of input proteins were used for Co-IP. (C) BiFC analysis of PRL1 with DCL1, HYL1, SE, AGO1 and CDC5. Paired cCFP- and nVenus-fusion proteins were co-infiltrated into <i>N. benthamiana</i> leaves. The BiFC signal (Yellow fluorescence) was detected at 48 h after infiltration by confocal microscopy, assigned as green color and marked with arrow. 30 nuclei were examined for each pair and an image is shown. Red: auto fluorescence of chlorophyll. (D) and (E) Co-immunoprecipitation between PRL1 and DCL1. (F) and (G) Co-immunoprecipitation between PRL1 and SE. PRL1-YFP or YFP were co-expressed with DCL1-MYC and SE-MYC in <i>N. benthamiana</i>, respectively. Anti-YFP and anti-MYC (MBL) antibodies were used to detect YFP- and MYC-fused proteins, respectively. The protein pairs in the protein extracts were indicated on the on tope of the picture and proteins detected by western blot were indicated on the left side of the picture. Ten percent of input proteins were used for IP and one percent of inputs proteins were used for Co-IP.</p

PRL1 is required for miRNA maturation <i>in vitro</i>.

<p>(A) and (B) A schematic diagram of the <i>MIR162b</i> (A) and <i>pre-miR162b</i> (B) used <i>in vitro</i> processing assay. (C) and (D) The amount of miR162b produced from <i>MIR162b</i> and <i>pre-miR162b</i> were reduced in <i>prl1-2</i>. Proteins were isolated from inflorescences of <i>prl1-2</i> and Col and incubated with <i>MIR162b</i> or <i>pre-miR162b</i>. The reactions were stopped at various time points as indicated in the picture. (E) and (F) Quantification of miR162b production in <i>prl1-2</i> compared to that in Col. Quantification analysis was performed at 80 min. The radioactive signal of miR162 were normalized to input and compared with that of Col. The amount of miR162 produced in Col was set as 1. The value represents mean of three repeats (*** <i>P</i><0.001; t-test).</p

The role of PRL1 in siRNA biogenesis.

<p>(A) PRL1 interacts with DCL3 and DCL4. Co-IP was performed to detect the interaction of PRL1 with DCL3 or DCL4. MBP and MBP-PRL1 fused protein were expressed in <i>E.coli</i>. YFP, DCL3-YFP and DCL4-YFP were expressed in <i>N. benthamiana</i> leaves. Anti-YFP was used for IP. For loading, ten percent and one percent of input proteins were used for IP and Co-IP, respectively. (B) <i>prl1-2</i> impairs siRNA production from double-stranded RNAs (dsRNAs). Protein extracts isolated from inflorescences of Col, <i>prl1-2</i> and <i>prl1-2</i> containing a PRL1-YFP transgene were incubated dsRNAs for 120 min. dsRNAs were synthesized through <i>in vitro</i> transcription of a DNA fragment (5′ portion of <i>UBQ5</i> gene, ∼460 bp) under the presence of [α-³²P] UTP.</p

Humidity-Driven Color-Fluorescence Dual Switching of Naphthalenediimide Aggregates

Numerous emerging applications require precise monitoring of humidity that is not only sensitive and miniaturized but also durable and portable. However, except for the existing sensing mechanisms, such as resistive, capacitive, impedance, and voltage-based devices, little attention has been paid to exploiting dual readout modes with novel properties that can meet the requirements of simplicity and convenience. In this work, an example of color/fluorescence dual-switching by using an aggregator building block of naphthalenediimide with a porous hygroscopic polymer matrix is proposed. With an increase in the water fraction, there are obvious variations in fluorescence, driven by H-aggregation of the aggregator, and in color, driven by intramolecular-charge transfer, which facilitate the monitoring of external humidity. More importantly, the prepared humidity-sensitive switch demonstrated excellent cycling stability and reversibility. This work may shed light on the precise fabrication of aggregation-activated materials for potential applications in humidity-sensitive systems and switching devices

Fully Integrated Ratiometric Fluorescence Enrichment Platform for High-Sensitivity POC Testing of Salivary Cancer Biomarkers

The point-of-care (POC) testing of cancer biomarkers in saliva with both high sensitivity and accuracy remains a serious challenge in modern clinical medicine. Herein, we develop a new fully integrated ratiometric fluorescence enrichment platform that utilizes acoustic radiation forces to enrich dual-emission sandwich immune complexes for a POC visual assay. As a result, the color signals from red and green fluorescence (capture probe and report probe, respectively) are enhanced by nearly 10 times, and colorimetric sensitivity is effectively improved. When illuminated using a portable UV lamp, the fluorescence color changing from red to green can be clearly seen with the naked eye, which allows a semiqualitative assessment of the carcinoembryonic antigen (CEA) level. In combination with a homemade smartphone-based portable device, cancer biomarkers like CEA are quantified, achieving a limit of detection as low as 0.012 ng/mL. We also directly quantify CEA in human saliva samples to investigate the reliability of this fully integrated platform, thus validating the usefulness of the proposed strategy for clinical diagnosis and home monitoring of physical conditions

Fully Integrated Ratiometric Fluorescence Enrichment Platform for High-Sensitivity POC Testing of Salivary Cancer Biomarkers

The point-of-care (POC) testing of cancer biomarkers in saliva with both high sensitivity and accuracy remains a serious challenge in modern clinical medicine. Herein, we develop a new fully integrated ratiometric fluorescence enrichment platform that utilizes acoustic radiation forces to enrich dual-emission sandwich immune complexes for a POC visual assay. As a result, the color signals from red and green fluorescence (capture probe and report probe, respectively) are enhanced by nearly 10 times, and colorimetric sensitivity is effectively improved. When illuminated using a portable UV lamp, the fluorescence color changing from red to green can be clearly seen with the naked eye, which allows a semiqualitative assessment of the carcinoembryonic antigen (CEA) level. In combination with a homemade smartphone-based portable device, cancer biomarkers like CEA are quantified, achieving a limit of detection as low as 0.012 ng/mL. We also directly quantify CEA in human saliva samples to investigate the reliability of this fully integrated platform, thus validating the usefulness of the proposed strategy for clinical diagnosis and home monitoring of physical conditions

Fully Integrated Ratiometric Fluorescence Enrichment Platform for High-Sensitivity POC Testing of Salivary Cancer Biomarkers

The point-of-care (POC) testing of cancer biomarkers in saliva with both high sensitivity and accuracy remains a serious challenge in modern clinical medicine. Herein, we develop a new fully integrated ratiometric fluorescence enrichment platform that utilizes acoustic radiation forces to enrich dual-emission sandwich immune complexes for a POC visual assay. As a result, the color signals from red and green fluorescence (capture probe and report probe, respectively) are enhanced by nearly 10 times, and colorimetric sensitivity is effectively improved. When illuminated using a portable UV lamp, the fluorescence color changing from red to green can be clearly seen with the naked eye, which allows a semiqualitative assessment of the carcinoembryonic antigen (CEA) level. In combination with a homemade smartphone-based portable device, cancer biomarkers like CEA are quantified, achieving a limit of detection as low as 0.012 ng/mL. We also directly quantify CEA in human saliva samples to investigate the reliability of this fully integrated platform, thus validating the usefulness of the proposed strategy for clinical diagnosis and home monitoring of physical conditions

Supplemental material for ⁶⁸Ga-somatostatin receptor analogs and ¹⁸F-FDG PET/CT in the localization of metastatic pheochromocytomas and paragangliomas with germline mutations: a meta-analysis

<p>Supplemental material for ⁶⁸Ga-somatostatin receptor analogs and ¹⁸F-FDG PET/CT in the localization of metastatic pheochromocytomas and paragangliomas with germline mutations: a meta-analysis by Ying Kan, Shuxin Zhang, Wei Wang, Jie Liu, Jigang Yang and Zhenchang Wang in Acta Radiologica</p

<i>urt1-3</i> increases the silique length in <i>hen1-2 heso1-2</i> and enhances miRNA function in <i>hen1-1 heso1-2</i>.

<p>(A) Fully-expanded siliques from plants of the indicated genotypes. Scale bar, 1cm. (B) Average silique length in indicated genotypes. 40 siliques from at least 6 individual plants for each genotype were analyzed. (C) Schematic structure of URT1 protein. PRO_Rich: Proline rich domain; GLN_Rich: Glutamine rich domain; PAP/25A: Poly A polymerase domain; Core: Core regions of PAP/25A; PAP/25A associated: Poly A polymerase associated domain. (D) Conservation of URT1 P618 across species. <i>At</i>, <i>A</i>. <i>thaliana</i>; <i>Hs</i>, <i>H</i>. <i>sapiens</i>; <i>Sp</i>, <i>S</i>. <i>pombe</i>; <i>Dm</i>, <i>D</i>. <i>melanogaster</i>; <i>Ce</i>, <i>C</i>.<i>elegans</i>; <i>Cr</i>, <i>C</i>. <i>reinhardtii</i>. (E) <i>urt1-3</i> enhances miRNA function in <i>hen1-1 heso1-2</i>. The transcript levels of five miRNA targets in indicated genotypes. Target mRNA accumulation in each genotype was quantified by qPCR using primers spanning the cleavage site and compared with those of wild type (Wt). The expression level of each gene in Wt was arbitrarily set to 1. Quantifications are normalized with <i>GAPDH</i> transcript. <i>ACT2</i> was served as an internal control. Values are means of three biological replicates ±SD.</p