Fig. 5

Expression level analysis of GTPBP2 protein in HeLa cells transfected with WT and mutant A536del GTPBP2 constructs. Cells were transfected with WT GTPBP2 or with A536del GTPBP2 cDNAs. Cells transfected with A536del GTPBP2 were then treated with the proteasome inhibitor MG132 (+). Untransfected cells were used as control. a) Total protein lysates from untransfected and transfected cells were obtained by solubilization. An equal quantity of protein was separated by SDS-PAGE and blotted onto nitrocellulose paper. The blots were incubated with polyclonal antibodies to GTPBP2. A cropped representative Western blot is shown. The lower panel is a Ponceau Red staining of the same cell lysates, used as loading control. b) Quantification of GTPBP2 protein bands was performed by densitometric analysis on western blots. Data (mean values from at least three independent experiments + S.D.) are reported as the percentage of values from WT GTPBP2 protein. Statistical analysis was performed by One-way ANOVA test, followed by multiple comparisons Dunnett’s test. * p < 0.05.