Fig. 1: The absence of neurofibromin decreases protein levels and enzymatic activity of respiratory complex I. | Cell Death & Differentiation

Fig. 1: The absence of neurofibromin decreases protein levels and enzymatic activity of respiratory complex I.

From: Tumor growth of neurofibromin-deficient cells is driven by decreased respiration and hampered by NAD+ and SIRT3

Fig. 1

OXPHOS protein levels were analyzed by Western immunoblot (WB, A) or blue native polyacrylamide gel electrophoresis (BN-PAGE, B). In A, calnexin was used as a loading control. In B, bands corresponding to different respiratory complexes were cut, run on an SDS-PAGE and probed for the expression of the complex I subunits NDUFB8, GRIM19, NDUFS1 and NDUFS3, of the complex V α subunit, of the complex III UQCRC2 subunit and of the complex II SDHB subunit. C Spectrophotometric analysis of the NADH dehydrogenase activity of complex I (CI) is shown as arbitrary units and normalized for citrate synthase (CS) activity. D BN-PAGE carried out on digitonized mitochondria to preserve assembled respiratory supercomplexes. Gels were either subjected to in gel complex I activity, or stained with Coomassie blue and transferred to PVDF membrane for protein identification. UQCRC1 and GRIM19 are complex III and complex I subunits, respectively. E Spectrophotometric analysis of the NADH dehydrogenase activity of complex I in control (empty vector, EV) and GRD Nf1−/− MEFs in basal condition (10 % FBS) or following growth in a media containing PDGF (10 ng/ml, and 0.5 % FBS). All experiments in the Figure were carried out in Nf1+/+ and Nf1−/− MEFs. Data are reported as mean ± SD values (n ≥ 3); ***p < 0.001; **p < 0.01; *p < 0.05 with a Student’s t test analysis.

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