Fig. 2: Hypoxic stress increases the expression of IR-A isoform.

A The Gene expression data for HIF-1α expression were extracted from Schafer-Welle (Scafer-Welle-56-MAS5.0-u133a) dataset using R2: Genomics Analysis and Visualization platform for 26 normal muscle samples and 30 RMS samples³⁵. The data distribution is represented by a box plot, the line inside the box represents the median and the whiskers represent minimum and maximum expression. p = 1.5e−28. B RT-PCR depicting the IR alternative splicing in HeLa S3 cells after 48 h of normoxia or hypoxia (1% O₂). Western blot analysis of the Hif-1α induction by hypoxia treatment is shown with β-actin as control. C Quantification of IR-B expression in control and hypoxic conditions; n = 6, p = 0.0018. D RT-PCR analysis for the INSR alternative splicing of the siRNA-mediated knockdown of Hif-1α followed by treatment of normoxia or hypoxia (1% O₂). Western blot analysis for Hif-1α expression and β-tubulin as a loading control. E Quantification of IR-B expression in control and hypoxic conditions of the Hif-1α knockdown experiments. n = 6, p = 0.0013. F Depiction of the wild-type insulin receptor (IR) minigene or the CUG-BP1 binding-site mutated minigene. RT-PCR depicting both INSR minigenes alternative splicing. G Quantification of the 10.11.12 isoform expression is shown n = 4, p = 0.405. Results are shown as the standard error of the mean (±SEM).