goat anti platelet derived growth factor receptor beta  (R&D Systems)

Journal: International Journal of Molecular Sciences

Article Title: Expression of Lumican and Osteopontin in Perivascular Areas of the Glioblastoma Peritumoral Niche and Its Value for Prognosis

doi: 10.3390/ijms26010192

Figure Lengend Snippet: CMA activity in PCs correlates with patient survival. ( A ) Representative images of peritumoral areas according to patient classification showing co-localization of puncta pattern expression of LAMP-2A protein (dark brown) with the PC marker α-SMA (pink) in microvessels of GB patients. Samples were classified as severe (highest α-SMA/LAMP-2A co-localization), moderate or mild (basal co-localization) according to the histological evaluation. A1 to A3 shows magnifications of microvessels (v). LAMP-2A co-localizationwith α-SMA + cells is marked with arrows. Scale bar: 100 µm. ( B ) Representative images of PCs marked with PDGFRβ (in red) in microvessels (v) showing co-localization of puncta pattern expression of LAMP-2A protein (in green) in peritumoral areas of severity classified GB patients. B1 to B3 show magnifications of PDGFRβ + cells. Positive co-localization (in yellow) is marked with arrows. Nuclei were stained with DAPI (blue). Scale bar: 100 µm. ( C ) Overall survival shown by Kaplan–Meier curves of the severity classification related to CMA activity in PCs; * p < 0.05: difference between mild and severe; # p < 0.05: difference between mild and moderate. ( D ) Age distribution by gender of the cohort of GB patients; **** p < 0.0001; ns indicates no significance. ( E ) Severity classification related to gender in the age range 30–65 years in the cohort of GB patients; **** p < 0.0001; ns indicates no significance.

Article Snippet: For fluorescent double-labeling of patient’s samples, goat anti-platelet-derived growth factor receptor beta (PDGFRβ; 1/250; BAF1042, R&D Systems, Barcelona, Spain), rabbit anti-LAMP-2A (1/1000; 51-2200, Invitrogen, Waltham, MA, USA) and mouse anti-RGS5 (1/150; MA5-25584, Invitrogen, Waltham, MA, USA) antibodies were used.

Techniques: Activity Assay, Expressing, Marker, Staining

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