Figure 4

Autophagy differences correlate with initial but not established tumorigenic activity. (A) Comparison of the effect of BafA₁ on LC3B-II levels (relative to ACTIN) in control- and KRAS^G12D-transduced human BCs or LPs cells assessed after another three days in vitro. Cells were incubated with 200 nM BafA1 for 2 h and then protein levels determined by WB analysis. Results shown are the mean ± SEM from experiments with cells from three normal individuals (BCs or LPs). (B) Representative views of immunostained LC3B in normal donor tissue and matching KRAS^G12D-induced de novo tumors. (C) Representative pictures of bioluminescent signals measured in mice 2 weeks after the mice were injected subcutaneously with KRAS^G12D-transduced G⁺ or R⁺ cells (left panels). The right panel shows the in vivo signals measured in all mice with the data for the G⁺ cells normalized to the matching R⁺ cell data (data for BC-derived cells shown in blue, n = 3, and for LP-derived cells in red, n = 5). P values were calculated using the paired Student t-test. Images were taken with Xenogen IVIS Lumina system and analyzed with Living Image version 3.0 software. (D) Similar bioluminescence data for secondary transplants of cells from 5 of the primary transplants of KRAS^G12D-transduced cells shown in (C).