Figure 3.

Effects of IGF1R localization over cell metabolism. (A) U87Empty, U87WT, and U87Mut cells were cultured 24 h in serum-free medium or with 50-nM IGF1 stimulation (IGF1). To test the specificity of the response, pre-incubation (1 h) with 0.5 μM OSI906 was also performed (IGF1+ OSI). (A, B) GLUT1 mRNA expression was calculated by rqPCR by the relative quantitation method. (A) Values are presented as fold change compared to control conditions (serum-free). (B) IGF1 stimuli comparison between cell lines. The results are presented as fold change due to IGF1 stimulation over basal conditions (serum-free). Values are expressed as mean ± SD of three independent experiments performed in triplicates (*p < 0.05, **p < 0.005, ***p < 0.001, ****p < 0.0001, ANOVA, Tukey’s post-test). (C) LDH enzyme activity was measured and normalized to DNA content (mg). The results are expressed as mean ± SD of three independent experiments (**p < 0.005, ANOVA, Tukey’s post-test). (D) Representative western blotting (n = 3) for U87Empty, U87WT, and U87Mut cell protein extracts. The membranes were blotted with anti-pPDC (line 1), anti-FAS (line 3), anti-ACSL5 (line 5), and anti-pS6K (line 7). Each blot was stripped and reprobed with anti-β actin (lines 2, 4, and 6) or antitotal-S6K (line 8). The relative quantification of the bands is shown under each line.