FIGURE 5.

Reconstitution of human Tid1 in mTid1-depleted NIH3T3 cells restores sensitivity of cells to rapamycin or EBSS-induced autophagy. A, NIH3T3 cells stably expressing NS shRNA or mTid1 shRNA were treated with 0.01, 0.1, or 1 μm rapamycin (Rapa) overnight. The levels of LC3-II and other related proteins as indicated were detected with specific antibodies. B, NIH3T3 cells stably expressing NS shRNA or mTid1 shRNA were incubated with EBSS for indicated time points, and the LC3-II levels were examined. C and D, mTid1-depleted NIH3T3 cells were reconstituted with human Tid1L (C), with human Tid1S (D), or control (Ctrl), and these cells were treated with rapamycin. The levels of LC3-II, Tid1L, or Tid1S were examined with anti-LC3 or anti-Tid1 antibodies.