Fig. 5
TBC1D2b hinders Rab22-mediated invasion. a Doxycycline-inducible murine TripZ-TBC1D2b construct was expressed in 344SQ and H1299 cell lines. Western blot demonstrates TBC1D2b expression over 48 h of time course induction. b Co-immunoprecipitation validates TBC1D2b interacts with Rab22 in 344SQ and 531LN2, however, no association was detected with Rab31 in these murine NSCLC cell lines (5% indicates whole cell lysate). c Quantification of Boyden chamber assay indicates 24 h of TBC1D2b overexpression diminishes cell migratory and invasive potential in 344SQ cell line; all asterisks indicate statistical significance by t-test (n ≥ 3, *p ≤ 0.05); error bars represent standard error mean. d Western blot confirms overexpression of human GFP-Rab22 in 393 P and H358 cell lines. e 393P_GFP-Rab22 overexpressing cells exhibit increased Boyden chamber migration and invasion; all asterisks indicate statistical significance by t-test (n ≥ 3, *p ≤ 0.05); error bars represent standard error mean. f pLenti-TBC1D2b construct was transiently expressed in 393P_GFP or 393P_GFP-Rab22 cell lines for 24 h prior to 24 h of doxycycline induction. mRNA confirms overexpression of TBC1D2b (48 h) and Rab22 overexpression (24 h). g In vitro transwell migration and invasion assay following TBC1D2b overexpression in 393P_GFP and 393P_GFP-Rab22 cell lines
