Figure 5: SRC kinases are activated by TNFα and BMP6, and can regulate Notch. | Nature Communications

Figure 5: SRC kinases are activated by TNFα and BMP6, and can regulate Notch.

From: TNFα drives pulmonary arterial hypertension by suppressing the BMP type-II receptor and altering NOTCH signalling

Figure 5

(a) Schematic depicting SRC phosphorylation. (b) Representative immunoblots of phospho-SRC Tyr527, Tyr416 and total SRC expression in human dPASMCs from disease-free controls and HPAH patients treated with TNFα (1 ng ml−1) and/or BMP2 (10 ng ml−1) or BMP6 (10 ng ml−1) for 30 min as indicated. Reprobed for α-tubulin to ensure equal loading. The arrows indicate the SRC bands. The data shown are representative of three control and HPAH cell lines. (c) Representative immunoblots of phospho-SRC Tyr527, Tyr416 and total SRC expression in human control dPASMCs following transfection with DharmaFECT1 alone (DH1), siBMPR2, or non-targeting siRNA control (siCP) and treated with TNFα (1 ng ml−1) and/or BMP6 (10 ng ml−1) for 30 min as indicated. Reprobed for α-tubulin to ensure equal loading. The data shown are representative of three experiments. (d) Representative immunoblots of phospho-SRC Tyr527, Tyr416 and total SRC expression in human HPAH dPASMCs following transfection with DH1 alone, siACVR2A, or siCP and treatment with TNFα (1 ng ml−1) and/or BMP6 (10 ng ml−1) for 30 min as indicated. Reprobed for α-tubulin to ensure equal loading. The data shown are representative of three cell lines. (e,f) NOTCH2 and NOTCH3 mRNA expression in human HPAH dPASMCs following transfection with DH1 alone, siFYN, siYES, siSRC or siCP and treatment with TNFα (1 ng ml−1) or BMP6 (10 ng ml−1) for 1 h as indicated. Expression was normalized to ACTB (n=3). (g) Proliferation of human HPAH dPASMCs on day 6 following transfection with DH1 alone, siFYN, siYES, siSRC or siCP and treatment every 48 h with TNFα (1 ng ml−1) and/or BMP6 (50 ng ml−1) as indicated (n=3). (h) Proliferation of human control dPASMCs on day 6 following transfection with DH1 alone, siFYN, siYES, siSRC or siCP and treatment every 48 h with TNFα (1 ng ml−1) and/or BMP6 (50 ng ml−1) as indicated (n=3). One-way analysis of variance with post hoc Tukey’s for multiple comparisons used in e,f,g and h. *P≤0.05, **P≤0.01, ***P≤0.001. Error bars represent mean±s.e.m.

Back to article page