mouse pdgf r alpha antibody  (Bio-Techne corporation)

Journal: PLOS ONE

Article Title: Colonic dysmotility regulated by downregulation of PDGFRα + cells / SK3 channel in DSS-induced colitis mice

doi: 10.1371/journal.pone.0312413

Figure Lengend Snippet: A–D. Western blot analysis of PDGFRα and SK3 in control and DSS-induced colitis mice. The data were analysed using densitometric quantification (% tubulin and normalized to data from control mice; n = 7, * P < 0.05). E-F. Quantitative RT‒PCR analysis of PDGFRα and SK3 expression in the colonic muscle layers of control and DSS-induced colitis mice. G. SK3 channels colocalized with PDGFRα + cells in the smooth muscle layer in the control and DSS-induced colitis mouse groups. The data were normalized to gapdh and the data from control mice ( pdgfr α n = 7; kcnn3 n = 8; * P < 0.05).

Article Snippet: The samples were incubated in 0.1 M phosphate-buffered saline (PBS) containing 10% normal goat serum for 2 h at 24°C to block nonspecific binding and incubated with a goat anti-PDGFRα antibody (1:200; AF1062, R&D Systems, USA) and a rabbit anti-SK3 (Ki67) antibody (1:50; GB13030, Wuhan Goodbio Technology, China) mixed with Triton-X100 (0.5%, Sigma–Aldrich, St. Louis, MO, USA) at 4°C for 24 h. The samples were washed with 0.1 M PBS for 30 min and then incubated at 24°C with Cy3-conjugated anti-goat IgG (1:300; GB21404, Wuhan Goodbio Technology, China), Alexa Fluor 488-conjugated goat anti-rabbit IgG (1:100, Jackson ImmunoResearch, USA) and DAPI for 2 h. Images were acquired using a confocal laser-scanning microscope (Leica TCS SP8, Germany).

Techniques: Western Blot, Control, Expressing

Journal: bioRxiv

Article Title: Intramuscular adipose tissue physically restricts functional muscle recovery

doi: 10.1101/2024.12.17.628009

Figure Lengend Snippet: A) Experimental design. B) Immunofluorescence of cross-sections of tibialis anterior (TA) muscles from WT and FAP no Pparγ female mice 7 days post GLY injury stained for fibro-adipogenic progenitors (FAPs; PDGFRα, magenta) and nuclei (visualized through DAPI; white). Yellow arrowheads indicate FAPs. Scale bar: 100µm. Quantification of total FAPs per 20x field in WT (n=8-14 TAs) and FAP no Pparγ (n= 8-14 TAs) female mice at 3-, 5-, 7- and 21-days post GLY injury. C) Immunofluorescence of TAs of WT and FAP no Pparγ female mice 3 days post GLY injury, stained for FAPs (PDGFRa, magenta) and the proliferation marker, Ki67 (green). Yellow arrowheads indicate proliferating FAPs. Scale bar: 100µm. Quantification of percent of proliferating FAPs at 3- and 5-days post injury of WT (n=8 TAs) and FAP no Pparγ (n=8 TAs). D) Immunofluorescence of cross-sections of TAs of WT and FAP no Pparγ female mice 5 days post GLY injury, stained for FAPs (PDGFRa, magenta) and the canonical apoptosis marker, cleaved caspase 3 (cC3, green) 5 days post injury. Yellow arrowheads indicate FAPs that are undergoing apoptosis. Scale bar: 100µm. Quantification of percent of FAPs undergoing apoptosis in WT (n= 7-11 TAs) and FAP no Pparγ (n= 8-13 TAs) mice 5- and 7-days post GLY injury. All data are represented as mean ± SEM. A multiple unpaired two-tailed t test followed by a Holm-Šídák post hoc test was used.

Article Snippet: Primary antibodies used were rabbit anti-Perilipin (1:1000; Cell Signaling, 9349 S), rabbit anti-MyoG (1:250; Proteintech Group 14688-1-AP), rabbit anti-cleaved Caspse 3 (1:500, Millipore Sigma AB3623), and goat anti-PDGFRα (1:250, R&D Systems #AF1062), rabbit anti-Ki67 (Abcam ab15580), chicken anti-GFP (1:1000, Avis lab).

Techniques: Immunofluorescence, Muscles, Staining, Marker, Two Tailed Test

Journal: eLife

Article Title: PDGFRα signaling regulates Srsf3 transcript binding to affect PI3K signaling and endosomal trafficking

doi: 10.7554/eLife.98531

Figure Lengend Snippet: ( A, B ) Scatter dot plots depicting average size of Rab5 puncta per cell ( A ) and Pearson’s correlation coefficient of signals from anti-Rab5 and anti-PDGFRα antibodies ( B ) in scramble and shSrsf3 cell lines in the absence or presence (15–60 min) of PDGF-AA stimulation. Data are mean ± s.e.m. *p<0.05 (two-way ANOVA followed by uncorrected Fisher’s LSD test). Shaded shapes correspond to independent experiments. Summary statistics from biological replicates consisting of independent experiments (large shapes) are superimposed on top of data from all cells; n = 20 technical replicates across each of three biological replicates. ( C–H”’ ) PDGFRα antibody signal (white or magenta) and Rab5 antibody signal (white or green) as assessed by immunofluorescence analysis of scramble and shSrsf3 cells in the absence or presence (15–60 min) of PDGF-AA stimulation. Nuclei were stained with DAPI (blue). White arrows denote regions of colocalization, which are expanded in (C’’’–H’’’). Scale bars: 20 μm ( C–H’’ ), 3 μm ( C’’’–H’’’ ). ( I ) Western blot (WB) analysis of whole-cell lysates (WCL) from scramble (left) and shSrsf3 (right) cell lines following a time course of PDGF-AA stimulation from 15 min to 4 hr, with anti-phospho-(p)-Akt and anti-Akt antibodies. Line graph depicting quantification of band intensities from n = 4 biological replicates as above. Data are mean ± s.e.m. *p<0.05; **p<0.01 (two-tailed, ratio paired t -test within each cell line and a two-tailed, unpaired t -test with Welch’s correction between each cell line). ( J ) Model of experimental results in which PI3K/Akt-mediated PDGFRα signaling results in the nuclear translocation of Srsf3 and the subsequent AS of transcripts to decrease levels of proteins that promote PDGFRα trafficking out of early endosomes. Figure 6—source data 1. Phospho-Akt western blots. Figure 6—source data 2. Akt western blots. Figure 6—source data 3. Relative induction values of phospho-Akt.

Article Snippet: The following antibodies were used for immunofluorescence analysis: Rab5 (1:200, C8B1, 3547, Cell Signaling Technology Inc), PDGFRα (1:20, AF1062, R&D Systems).

Techniques: Immunofluorescence, Staining, Western Blot, Two Tailed Test, Translocation Assay

Journal: eLife

Article Title: PDGFRα signaling regulates Srsf3 transcript binding to affect PI3K signaling and endosomal trafficking

doi: 10.7554/eLife.98531

Figure Lengend Snippet:

Article Snippet: The following antibodies were used for immunofluorescence analysis: Rab5 (1:200, C8B1, 3547, Cell Signaling Technology Inc), PDGFRα (1:20, AF1062, R&D Systems).

Techniques: shRNA, Recombinant, Plasmid Preparation, Sequencing, Software, Microscopy