Figure 6. Arc deficiency increases Bax expression and cell death.

A, Western blot for Bak and Bax from quadriceps lysates of Nol3^-/-, Sgcd^-/-, and Nol3^-/-Sgcd^-/- mice. (β-tubulin serves as a loading control). B, Western blot for Bax and Bcl-X_L from mitochondrial protein fractions isolated from pooled hindlimb muscles of WT, Nol3^-/-, Sgcd^-/-, and Nol3^-/-Sgcd^-/- mice. (voltage-dependent anion channel (VDAC) serves as a mitochondrial protein loading control). C, Western blotting for Arc from lysates derived from WT and Bak^-/-Bak1^-/- SV40 transformed MEFs. Skeletal muscle lysates were included to show the enrichment of Arc in terminally differentiated cell types, while GAPDH serves as a protein loading control. D, Western blot for Bax and Arc in SV40 transformed MEFs infected with lentivirus expressing scrambled shRNA (con) or 3 different Bax-directed shRNAs. (GAPDH serves as a loading control). E, Western blot for Arc in SV40 transformed MEFs infected with lentivirus expressing either a scrambled shRNA or one of the Bax shRNAs and treated with proteosomal inhibitors Bortezomib or MG-132. WT MEFs are a control for normal endogenous Arc expression and GAPDH serves as a loading control. F, Western blot for Arc in SV40 transformed MEFs infected with a lentivirus expressing either a scrambled shRNA (con.) or shRNA directed against Arc. Western blots presented are quantified and statistically analyzed in Figure S3A-G. G, Quantification of dead cells by flow cytometry sorting for Annexin and PI positivity in the experimental groups shown, treated or untreated with staurosporin for 12 hours. *P<0.05 vs WT untreated. Experiment was run in triplicate.