pp1  (MedChemExpress)

Journal: BMC Molecular and Cell Biology

Article Title: Fyn kinase regulates dopaminergic neuronal apoptosis in animal and cell models of high glucose (HG) treatment

doi: 10.1186/s12860-021-00398-y

Figure Lengend Snippet: Inhibition of Fyn alleviates HG-induced cell injury by suppressing the mTOR/S6K signaling pathway. (A) Elevated cleaved caspase 3 levels induced by HG were rescued by rapamycin (100 nM) or PP1 (10 μM) treatment. PP1 inhibited p-Y416, but rapamycin did not change p-Y416 levels ( n = 5). β-actin, cleaved caspase 3 and Fyn were cropped from different parts of the same gel, and p-Y416 was cropped from a different gel. (B) PP1 or rapamycin inhibited mTOR/S6K ( n = 5). β-actin and S6K were cropped from different parts of the same gel, and p-S6K, mTOR and p-mTOR were cropped from different gels. (C) Flow cytometry showed that rapamycin or PP1 decreased the elevated cell apoptotic death induced by HG ( n = 3). (D) Decreased cell viability in response to HG was rescued by rapamycin or PP1 administration (n = 5)

Article Snippet: Anti-tyrosine hydroxylase (TH) (ab112, Abcam, Cambridge, UK); Anti-TH (sc-25,269, Santa Cruz, Santa Cruz, USA); anti-cleaved Caspase 3 (9664; CST, Danvers, USA); Anti-Fyn (ab184276, Abcam, Cambridge, UK); anti-p-Y416 (6943; CST, Danvers, USA); anti-mTOR (2972; CST, Danvers, USA), anti-p-mTOR (Ser2448) (5536; CST, Danvers, USA); anti-p70 S6 Kinase (9202; CST, Danvers, USA); anti-p-S6K(Thr389) (97,596; CST, Danvers, USA); anti-β-actin (20536–1-AP, Proteintech, Wuhan, China), FITC-conjugated goat anti-rabbit IgG (Proteintech, Wuhan, China); FITC-conjugated donkey anti-mouse (Proteintech, Wuhan, China); Alexa Fluor 555-conjugated donkey anti-rabbit (Beyotime, Haimen, China); PP1 (HY-13084,MCE,New Jersey, USA); rapamycin (HY-10219, MCE, New Jersey, USA); all-trans retinoic acid (RA) (R2625, Sigma–Aldrich, Missouri, USA); glucose (15,023,021,Thermofisher, Waltham, USA).

Techniques: Inhibition, Flow Cytometry