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. 2017 Jul 7;31(11):4734–4744. doi: 10.1096/fj.201700197RR

Figure 4.

Figure 4.

Gβ5 knockout impairs M3R-stimulated ERK1/2 phosphorylation in MIN6 cells. A) Gnb5+/+ (control) or knockout (Gβ5-KO) cells were stimulated with 100 μM Oxo-M for the indicated periods (minutes) and subjected to Western blot analysis with anti-phospho-ERK1/2 and anti-actin antibodies. Shown is a representative immunoblot. B) Quantitative analysis of Western blots (n = 3). Mean ± sd. At least 5 additional experiments with individual time points had a similar reduction of ERK1/2 phosphorylation in Gβ5-KO (data not shown). C) The control and knockout cells were stimulated with Oxo-M in the presence of inhibitors of protein kinases PKC (BIM, bisindolylmaleimide I), Src (GW5074 and PP2), EGFR (CL-387785), Erk1/2 (FR180204), MKK1/2 (U0126), and sorafenib (Raf and other Tyr-kinases), and insulin secretion was measured. D) The Gβ5-knockout and control MIN6 cells were stimulated with Oxo-M in the presence or absence of 100 ng/ml PMA, which was added with Oxo-M. E) Cells were treated as in D with 50 ng/ml EGF. Data in C–E are means ± sd from 5 independent experiments. *P < 0.05; **P < 0.01; ***P < 0.001.