Figure 2
Inceptor is highest in neurons and immunoprecipitated with IR and IGF1R in cortex, but may not significantly change Insulin/IGF signaling following knockdown. (a) Inceptor mRNA transcripts measured by cell type using qPCR and quantified relative to tata-binding-protein (TBP) as a control. Cells were cultured from C57BL/6 J mice or MIN6 immortalized line. (b) Blots for Inceptor CO-IP, the related IP (IR or IGF1R) plus input sample blots from the pancreas, liver, cortex, and hypothalamus from experiment performed on two animals. (c) Knockdown of Inceptor with lentiviral construct #3. Total inceptor knockdown is shown via western blot compared to actin (on one western blot), and a second blot of the same samples shows total levels of pERK, ERK, pAKT, and AKT. Each lane within a group represents a biological replicate (n of 3–4). (d) Statistical analysis of western blots. Statistical significance determined with unpaired t-test for Inceptor knockdown, and two-way ANOVA with Tukey for multiple comparisons for pERK/ERK and pAKT/AKT. There was no statistically significant difference in the interaction term (insulin treatment X inceptor knockdown), p = 0.1346. Some blots here and in other figures have been contrast and brightness adjusted for maximum visibility. Full raw images of all blots are available as noted in Data Availability section, below.
