Additional file 1 of Hsa_circ_0022383 promote non-small cell lung cancer tumorigenesis through regulating the miR-495-3p/KPNA2 axis

Supplementary Material

Data for: Diurnal Variations of Greenhouse Gases Emissions from Coastal Reclamation Marine Aquaculture Ponds

Ranges and means of physicochemical parameters in the coastal reclamation marine culture ponds of Tianjin. CO2, CH4, N2O concentration and saturation in water of the DG and HG ponds. CO2, CH4, N2O emission fluxes at water-air interface of the DG and HG ponds

Additional file 1 of Role of Secretoglobin+ (club cell) NFκB/RelA-TGFβ signaling in aero-allergen-induced epithelial plasticity and subepithelial myofibroblast transdifferentiation

Additional file 1: Table S1. PCR Primers. Figure S1. Percentage of live cells from BAL fluid of mice treated for 4 days with CDE, n=6-8. Figure S2. C57BL/6J mice were treated with daily challenges of CDE or PBS i.n. over 4 d. Immunofluorescence microscopy (IFM). Sections were stained with (a) anti-TGFβ1, (b) anti-RelA, and (c) anti-Snail antibody (Ab, red color) and counter-stained with DAPI (blue) to visualize nuclei; n = 6-8 animals/group. Figure S3. Single cell flow cytometry using a 4-day CDE exposure model. Representative gating for cell selection using Fluorescence Minus One (FMO) controls to select the correct gate for SMA+ and CD326+ CDE-exposed cells; n=5. Figure S4. C57BL/6J mice were treated with daily challenges of CDE or PBS i.n. and given BMS-345541 (IKKi) i.p. simultaneously over 5 d, n=5-6. A. IFM. Sections were stained with anti p-SMAD3 Ab (red) and DAPI (blue). Figure S5. Scgb1a1CreERTM/+ × RelAfl/fl mice were pre-treated with TX for 10 d and challenged with/without CDE over 4 d, n=5-8. A. IFC of dual stain between α-SMA (green) and Collagen I (purplered) with blue DAPI blue nuclear staining B. IFC Sections were stained with anti-TGFβ1 Ab (red) and counterstained with DAPI (blue). Images obtained at 20X

Data for: Diurnal Variations of Greenhouse Gases Emissions from Coastal Reclamation Marine Aquaculture Ponds

Ranges and means of physicochemical parameters in the coastal reclamation marine culture ponds of Tianjin. CO2, CH4, N2O concentration and saturation in water of the DG and HG ponds. CO2, CH4, N2O emission fluxes at water-air interface of the DG and HG ponds

Data for: Diurnal Variations of Greenhouse Gases Emissions from Coastal Reclamation Marine Aquaculture Ponds

Ranges and means of physicochemical parameters in the coastal reclamation marine culture ponds of Tianjin. CO2, CH4, N2O concentration and saturation in water of the DG and HG ponds. CO2, CH4, N2O emission fluxes at water-air interface of the DG and HG ponds

Data for: Diurnal Variations of Greenhouse Gases Emissions from Coastal Reclamation Marine Aquaculture Ponds

Ranges and means of physicochemical parameters in the coastal reclamation marine culture ponds of Tianjin. CO2, CH4, N2O concentration and saturation in water of the DG and HG ponds. CO2, CH4, N2O emission fluxes at water-air interface of the DG and HG ponds

Downregulation of MRP1 decreased MDR1 expression in MC3/5FU cells.

<p>MC3/5FU cells were transfected with plasmids containing a ABCC1/MRP1 specific shRNA and a non-specific control shRNA, the resulting clones were MC3/5FU–S (S for silenced) and MC3/5FU-NS (NS for non-silenced). Total RNA and total protein were extracted from the 2 cell lines and then subjected to RT-PCR and Western blot respectively. (A) Quantitative real-time RT–PCR result (n = 3). The values were nomalized with GAPDH. When ABCC1/MRP1 mRNA was significantly down-regulated in MC3/5FU-S cell, ABCB1/MDR1 mRNA was simultaneously significantly down-regulated. (B) Representative RT-PCR results (n = 3). GAPDH was used as loading control. (C) Representative Western-blot results. β-actin was used as loading control. ** P<0.01 vs MC3/5FU-S, *** P<0.001 vs MC3/5FU-S.</p

Strong positive correlation was found between MDR1 and MRP1/nuclear MRP1 expression in MEC tissues.

<p>Immunohistochemistry (IHC) staining was conducted to determine the localization and expression of MRP1 in formalin-fixed, paraffin-embedded mucoepidermoid carcinoma tissues using anti-MRP1 antibody. Results were scored in qualitative and quantitative way. The tissues were counterstained with hematoxylin. Brownish staining indicated MRP1 (a, b, c) or MDR1 (d, e, f) expression in the tissues. In the strongly MRP1/nuclear MRP1 stained MEC tissues (a), MDR1 was also strongly expressed (d). And MDR1 was higher expressed in cells with higher nuclear MRP1 expression compared with cells with lower nuclear MRP1 expression. In the moderately MRP1 expressed MEC tissues (b), moderately expressed MDR1 (e) were found both in the cytoplasm and cytomembrane of MEC cells. In the weakly MRP1 expressed MEC tissues (c), faint expression of MDR1 (f) was detected on the cytomembrane of MEC cells (200×magnification).</p

Nuclear Multidrug-Resistance Related Protein 1 Contributes to Multidrug-Resistance of Mucoepidermoid Carcinoma Mainly via Regulating Multidrug-Resistance Protein 1: A Human Mucoepidermoid Carcinoma Cells Model and Spearman's Rank Correlation Analysis

<div><p>Background</p><p>Multidrug resistance-related protein 1 (MRP1/ABCC1) and multidrug resistance protein 1 (MDR1/P-glycoprotein/ABCB1) are both membrane-bound drug transporters. In contrast to MDR1, MRP1 also transports glutathione (GSH) and drugs conjugated to GSH. Due to its extraordinary transport properties, MRP1/ABCC1 contributes to several physiological functions and pathophysiological incidents. We previously found that nuclear translocation of MRP1 contributes to multidrug-resistance (MDR) of mucoepidermoid carcinoma (MEC). The present study investigated how MRP1 contributes to MDR in the nuclei of MEC cells.</p><p>Methods</p><p>Western blot and RT-PCR was carried out to investigate the change of multidrug-resistance protein 1 (MDR1) in MC3/5FU cells after MRP1 was downregulated through RNA interference (RNAi). Immunohistochemistry (IHC) staining of 127 cases of MEC tissues was scored with the expression index (EI). The EI of MDR1 and MRP1 (or nuclear MRP1) was analyzed with Spearman's rank correlation analysis. Using multiple tumor tissue assays, the location of MRP1 in other tissues was checked by HIC. Luciferase reporter assays of MDR1 promoter was carried out to check the connection between MRP1 and MDR1 promoter.</p><p>Results</p><p>MRP1 downregulation led to a decreased MDR1 expression in MC3/5FU cells which was caused by decreased activity of MDR1 promoter. IHC study of 127 cases of MEC tissues demonstrated a strong positive correlation between nuclear MRP1 expression and MDR1 expression. Furthermore, IHC study of multiple tumor tissue array sections showed that although nuclear MRP1 widely existed in MEC tissues, it was not found in normal tissues or other tumor tissues.</p><p>Conclusions</p><p>Our findings indicate that nuclear MRP1 contributes to MDR mainly through regulating MDR1 expression in MEC. And the unique location of MRP1 made it an available target in identifying MEC from other tumors.</p></div

Nuclear MRP1 was a widespread phenomenon in MEC tissues.

<p>To exclude the possibility that MRP1 was expressed on the surface of nuclear membrane of MEC cells, the location of MRP1 was layer scanned by immunofluorescent confocal laser-microscopy. (A) 5×10⁴ MEC-1 cells were cultured on glass coverslips for 24 hours and then subjected to immunofluorescent cytochemistry staining. The cells were reacted with mouse monoclonal antibody specific for human MRP1, and were then stained with DyLight™594-conjugated AffinPure Dongkey Anti-Mouse IgG (b) and counterstained with DAPI (a) to reveal nuclei. (c) Merged pictures. The merged pictures showed that MRP1 was localized on the cytomembrane and the nuclei of MEC-1 cells which were derived from MEC tissue whose MRP1 expression was negligible in the immunohistochemistry (IHC) study. (B) MEC tissue with obvious nuclear MRP1 expression by IHC was subjected to immunofluorescent histochemical staining. MRP1 were reacted with mouse monoclonal antibody specific for human MRP1, then stained with Fluorescein-Conjugated AffiniPure Goat Anti-Mouse IgG second antibody (b) and counterstained with DAPI (a) to reveal nuclei, (c) phase contrast picture of MEC tissue, (d) merged picture. The merged picture showed that MRP1 was mainly localized in the nuclei of cells of MEC tissues.</p