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. 2016 Aug 16;6:31222. doi: 10.1038/srep31222

Figure 2. Insulin enhances SRSF1 expression and INSR exon 11 inclusion through the MAPK-ERK pathway.

Figure 2

(A) MIN6 cells were starved for 24 hours in 0.1% serum and then stimulated with 100 nM insulin in the presence or absence of 10 μM U0126 or vehicle (0.025% DMSO). Total RNA was extracted and subjected to RT-PCR using primers from exon 10 and 12 of INSR. GAPDH was used as a control. (B,C) Mouse and human pancreatic islets were starved for 24 hours in 0.1% serum and then stimulated with 100 nM insulin for 24 hours, in the presence or absence of 10 μM U0126. RNA was extracted and INSR splicing detected as in (A). GAPDH served as a RT-PCR control. The right panels (A–C) are quantification of % exon 11 inclusion from 3 experiments. Error bars, SD; n = 3, *p < 0.05. (D) RNA was extracted from human islets described in (C) and subjected to qRT-PCR using primers specific for SRSF1. Tubulin was used for normalization. (E) RNA was extracted from MIN6 cells described in (A) and SRSF1 mRNA levels were detected by qRT-PCR using primers specific for SRSF1. Tubulin was used for normalization. (F) Western blot of protein lysates from cells described in (E).