Journal: Journal of molecular and cellular cardiology
Article Title: Endothelial SHP-1 regulates diabetes-induced abnormal collateral vessel formation and endothelial cell senescence.
doi: 10.1016/j.yjmcc.2025.03.005
Figure Lengend Snippet: Fig. 7. SHP-1 regulates endothelial senescence under HG level exposure. (A) Representative images of senescence-associated β-galactosidase assay and (B) quantification of the percent of positive cells (blue staining). Immunoblot representative image and quantification of (C) p21, (D) Nrf2, and (E-F) phospho-pp2aC (Y307). EC were infected with adenoviral GFP, SHP-1 dominant-negative and native form and exposed to normal glucose (NG; 5.6 mmol/L; white bars) or high glucose (HG; 25 mmol/L; black bars) under hypoxia (1 % O2) for the last 16 h of treatment as well as in (F) nondiabetic (white bars) and diabetic (black bars) mice without (ec-SHP-1+/+) or with endothelial-specific SHP-1 deletion (ec-SHP-1−/−). Results are shown as mean ± SD of N = 7 independent cell replicates or mice per group. One-way ANOVA with Tukey's post hoc test. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)
Article Snippet: Primary antibodies for immunoblotting and immunohistochemistry; Actin (HRP; Sc-1616), GAPDH (HRP; sc-20,357), p21 (Sc-397), p53 (sc6243) and p-PP2a Cα/β Y307 (sc-271,903) were purchased from Santa Cruz Biotechnology (Santa Cruz, CA); p-Akt (S473), Akt (9272), pVEGFR2 Y1175 (2478), VEGFR2 (2479), p-eNOS Ser1171 (9571), eNOS (32027), NRF2 (12721) and secondary antibody of anti-rabbit and antimouse peroxidase-conjugated from Cell Signaling (Beverly, MA); PP2a Cα (610555) from BD bioscience, SHP-1 (ab32559), α-smooth muscle actin (Ab5694) from Abcam (Cambrige, UK); CD31 (HS-351117) from HistoSure (Goettingen, Germany); and Alexa 594–conjugated anti-rat and 488-conjugated anti-rabbit from Jackson ImmunoResearch Laboratories (West Grove, PA).
Techniques: Staining, Western Blot, Infection, Dominant Negative Mutation
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