Fig. 7.

Certain mutations of ETFA and ETFB found in GA-II patients hinder the neddylation of these substrates. (A and B) IB analysis of the effects of MG132 on the expression of Myc-tagged (A) or GFP-tagged ETF proteins (B) with or without the indicated mutation found in certain GA-II patients. hETFA, human ETFA; hETFB, human ETFB; TM, T₂₆₆M; QH, Q₁₂₅H; DN, D₁₂₈N; Δ, Δ_73–125. Quantification is shown as numbers under each image, determined using ImageJ. (C and D) The neddylation of Myc-tagged (C) or GFP-tagged ETF proteins (D) with or without the indicated mutation was examined as described in Fig. 5I (C) or Fig. 5L (D), respectively. (E and F) The ubiquitination of Myc-tagged (E) or GFP-tagged ETF proteins (F) with or without the indicated mutation was examined as described in Fig. 6I. (G and H) Twenty-four hours after BNL CL.2 cells were transfected with GFP-tagged ETF expression constructs with or without the indicated mutation, the possible impact on ER stress (G) and the possible colocalization of these exogenous proteins with mitochondria (H) were examined by IF analysis with an antibody against GRP78 (G) or Tom20 (H). (Scale bars, 10 μm.)