BV treatment does not activate the autophagic signal pathway. NSC34 cells were transiently transfected with the GFP control vector or GFP-tagged wild-type hSOD1 or mutant hSOD1G85R. Approximately 24 hrs after transfection, the cells were treated with 2.5 μg/ml BV or equal volume of saline for 24 hrs or left untreated. The cells were lysed using RIPA buffer, and western blot analysis was performed with anti-GRP78, anti-LC3, and anti-ISG15 antibodies to examine the expression of autophagic-degradation proteins (A). The arrowhead indicates LC3II protein levels. The membranes were reprobed with an anti-α-tubulin antibody as a loading control. Quantification of LC3II (B) and ISG15 (C) are shown. The values shown are the means ± SEM of data obtained from three independent experiments. * p < 0.05, ** p < 0.01.