Figure 5

(A) miR-221-5p (miR-221*) expression in skin fibroblasts from patients with CS and related transgenic cell models (n = 3). (B) Description of the HRAS/miR-221*/AMPK pathway. (C and D) Rescue of AMPKα2 expression by anti–miR-221-5p at the protein and the mRNA levels, respectively (n = 3). (E) Rescue of AMPKα2 expression, T172_P-AMPK/total AMPK ratio, and PGC1α expression by the anti–miR-221-5p (n ≥ 3). (F) Rescue of mitochondrial respiration by anti–miR-221-5p and its inhibition by a miR-221-5p mimic (n ≥ 3). (G) Rescue of mitochondrial steady-state ATP content by the anti–miR-221-5p (n ≥ 3). (H and I) Rescue of mitochondrial particles number by anti–miR-221-5p (n = 3). Original magnification, ×1000. Two-way ANOVA with Dunnett’s correction for multiple testing was used to compare the 3 groups of cells expressing HRAS p.G12A, HRAS p.G12S, or the empty plasmid control (pV) treated with the anti-miR scramble or the anti–miR-221-5p. Unpaired t test was used to compare the 2 groups of cells obtained from patients with CS and controls. Data are expressed as the mean ± SEM.*P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001.