antibody rabbit polyclonal anti stim1 cell signaling 4916s rrid ab 1849882  (Cell Signaling Technology Inc)

Journal: Respiratory Research

Article Title: Silencing of STIM1 attenuates hypoxia-induced PASMCs proliferation via inhibition of the SOC/Ca 2+ /NFAT pathway

doi: 10.1186/1465-9921-14-2

Figure Lengend Snippet: The effects of chronic hypoxia on STIM1 mRNA and protein expression. Representative Western blot images ( A ) and summarized data ( B ) for STIM1 proteins in distal (>4th generation) intrapulmonary arteries from control rats (N) and rats exposed to hypobaric hypoxia for 21 days (CH). β-actin was used as a loading control. Graph shows protein expression of STIM1 normalized to an average level in N. The results are expressed as the mean ± SEM for five experiments *: P < 0.05 compared to the N group. ( C ) STIM1 (red) and α-smooth muscle actin (ACTA2) (green) immunofluorescence staining in small pulmonary arteries from N and CH. Nuclei are counterstained with DAPI (blue). Scale bar = 10 μm. The results shown are from a single experiment and are representative of three separate experiments. ( D ) Analysis of STIM1 mRNA expression in distal (>4th generation) intrapulmonary arteries from N and CH. Parallel amplification of the rat housekeeping β-actin gene was used as an internal control. The results are expressed as the mean ± SEM for five experiments. *: P < 0.05 compared to the N group.

Article Snippet: Monoclonal Abs against STIM1 (BD Bioscience), Lamin B1 (Santa Cruz) and β-actin (Santa Cruz) or a polyclonal antibody against NFATc3 (Santa Cruz) were used as primary Abs.

Techniques: Expressing, Western Blot, Immunofluorescence, Staining, Amplification

Journal: Respiratory Research

Article Title: Silencing of STIM1 attenuates hypoxia-induced PASMCs proliferation via inhibition of the SOC/Ca 2+ /NFAT pathway

doi: 10.1186/1465-9921-14-2

Figure Lengend Snippet: The effect of STIM1 silencing on rat PASMC proliferation and cell cycle progression under hypoxia. ( A ). Western blot showing STIM1 and β-actin protein expression from untreated control PASMCs (Control) and PASMCs treated with transfection-nontargeted siRNA (siNT) or siRNA targeted to STIM1 (siSTIM1). ( B ). Mean ratios of STIM1 protein expression relative to that of β-actin, as measured by Western blotting in Control, siNT, and siSTIM1 PASMCs. *: P < 0.05 compared to the Control group and the siNT group. The results are expressed as the mean ± SEM of four experiments. ( C )After transfection with STIM1 siRNA, PASMCs were cultured in hypoxic conditions for 24 hours. [3H]-thymidine (3H-TdR) incorporation was measured to detect PASMC proliferation. ( D ) PASMCs were harvested for flow cytometry-based cell cycle analysis. PASMCs were either left untreated under normoxic conditions (N), left untreated under hypoxic conditions (HCON), treated with nontargeted siRNA under hypoxic conditions (HsiNT), or treated with siRNA targeted to STIM1 under hypoxic conditions (HsiSTIM1). *: P < 0.05 compared with the Control group and #: P < 0.05 compared with the HCON and HsiNT group. The results are expressed as the mean ± SEM of three experiments.

Article Snippet: Monoclonal Abs against STIM1 (BD Bioscience), Lamin B1 (Santa Cruz) and β-actin (Santa Cruz) or a polyclonal antibody against NFATc3 (Santa Cruz) were used as primary Abs.

Techniques: Western Blot, Expressing, Transfection, Cell Culture, Flow Cytometry, Cell Cycle Assay

Journal: Respiratory Research

Article Title: Silencing of STIM1 attenuates hypoxia-induced PASMCs proliferation via inhibition of the SOC/Ca 2+ /NFAT pathway

doi: 10.1186/1465-9921-14-2

Figure Lengend Snippet: The effect of STIM1 silencing on hypoxia-induced enhancement of SOC-mediated Ca 2+ influx. After transfection with STIM1 siRNA, PASMCs were cultured in hypoxic conditions for 24 hours. The SOC-mediated PASMC Ca 2+ influx was measured following stimulation with 10 μM cyclopiazonic acid during the change from Ca 2+ -free conditions to 2.5 mM Ca 2+ . PASMCs were either left untreated under normoxic conditions (N), left untreated under hypoxic conditions (HCON), treated with nontargeted siRNA under hypoxic conditions (HsiNT), or treated with siRNA targeted to STIM1 under hypoxic conditions (HsiSTIM1). A : Representative raw trace illustrating the changes in [Ca 2+ ]i (presented as fluorescence intensity (FI) in PASMCs in different groups. B : Summary of the data illustrated in showing averaged changes in fluorescence after 2.5 mM Ca 2+ restoration. *: P < 0.05 compared with the Control group and #: P < 0.05 compared with the HCON and HsiNT group. The results are expressed as the mean ± SEM of three separate experiments.

Article Snippet: Monoclonal Abs against STIM1 (BD Bioscience), Lamin B1 (Santa Cruz) and β-actin (Santa Cruz) or a polyclonal antibody against NFATc3 (Santa Cruz) were used as primary Abs.

Techniques: Transfection, Cell Culture, Fluorescence

Journal: Respiratory Research

Article Title: Silencing of STIM1 attenuates hypoxia-induced PASMCs proliferation via inhibition of the SOC/Ca 2+ /NFAT pathway

doi: 10.1186/1465-9921-14-2

Figure Lengend Snippet: The effect of STIM1 silencing on hypoxia-induced nuclear translocation of NFATc3 in PASMCs. NFATc3 staining was visualised by confocal microscopy and immunofluorescence. The primary antibody against NFATc3 was detected using a FITC-conjugated AffiniPure goat anti-rabbit IgG secondary antibody (green). The slides were counterstained with the nuclear dye DAPI (blue). ( A ): Immunofluorescence image of NFATc3 in PASMCs: PASMCs were either left untreated under normoxic conditions (N), left untreated under hypoxic conditions (HCON), treated with nontargeted siRNA under hypoxic conditions (HsiNT), or treated with siRNA targeted to STIM1 under hypoxic conditions (HsiSTIM1). Representative Western blot images ( B ) and summarized data ( C ) for NFATc3 nuclear levels, Lamin B1 was used as a loading control. *: P < 0.05 compared with the Control group and #: P < 0.05 compared with the HCON and HsiNT group. The results are expressed as the mean ± SEM of three separate experiments.

Article Snippet: Monoclonal Abs against STIM1 (BD Bioscience), Lamin B1 (Santa Cruz) and β-actin (Santa Cruz) or a polyclonal antibody against NFATc3 (Santa Cruz) were used as primary Abs.

Techniques: Translocation Assay, Staining, Confocal Microscopy, Immunofluorescence, Western Blot

Journal: Developmental cell

Article Title: TAOK2 is an ER-localized kinase that catalyzes the dynamic tethering of ER to microtubules

doi: 10.1016/j.devcel.2021.11.015

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Mouse monoclonal anti-Stim1 , Santa Cruz Biotech , sc-166840; RRID: AB_2198006.

Techniques: Recombinant, Purification, Software