Figure 2

Interrupting the dynamics of regular microtubules with either nocodazole or paclitaxel impairs conversion of LC3I to LC3II only in mitotic cells. (A) Immunoblot of lysates of fractioned HeLa cells. Mitotic cells were enriched by synchronization with 2 mM thymidine for 20 hr followed by release from the thymidine blockade for 20 hr in the absence or presence of 10 μM paclitaxel or nocodazole as described previously [40]. NH4Cl (20 mM) was added simultaneously with microtubule interfering agents. Loosely attached mitotic cells (S) were harvested by vigorous shake off from the remaining mixture (A) of interphase cells and mitotic cells attached to the dish. (B) The relative intensities of LC3I and LC3II bands were calculated from the scanned intensities of bands of three independent experiments as exemplified by immunoblot shown in (A). **, p < 0.01 as calculated by Student's T-test.