Fig. 6
CBAP loss affects insulin-stimulated lysosomal translocation of TSC2. a Analysis of insulin-stimulated Akt-TSC2-p70S6K signaling activation in control and CBAP-KO Hela cells. Cells were serum starved for 16 h and then stimulated with insulin for 15 min prior to lysis. Western blots were performed using indicated antibodies. b–d Hela cells (wild type, CBAP-KO or HA-CBAP-reconstituted) were treated as in (a), prior to immunofluorescent co-labeling of TSC2 with LAMP2 (b), with LysoView (c), or with HA-CBAP plus LysoView (d). The data are representative of three independent biological replicates (n = 45~ 95 cells). The degree of co-localization between TSC2 and LAMP2 or LysoView is shown in the right panel. Box represents the median and interquartile range, whereas whiskers represent maximum and minimum values. For the co-localization images, representative magnified insets are shown in the rightmost panel (top: TSC2; middle: LAMP2 or LysoView; bottom: merged). e Relative lysosomal co-localization of TSC2 upon insulin stimulation. Statistical data were compiled from results in (b) (left panel), or (c) and (d) (right panel). Error bars denote ± SEM. *P < 0.05; **P < 0.01; ***P < 0.001, according to two-tailed unpaired Student's t tests
