Fig. 2

Acidity does not inactivate rapamycin. a HT29 cells were treated with rapamycin (100 nM) that was previously incubated in DMEM full medium buffered at pH 7.4 or 6.4 for 24 h. As a control HT29 cells were also treated with fresh rapamycin. After 3 h of treatment, cells were lysed and lysates analyzed by Western blot. b HT29 cells were infected with lentiviruses expressing a control or raptor shRNA. Following selection, cancer cells were lysed and Western blot analysis was performed with the indicated antibodies. c HT29 cells generated in panel b were exposed to various pH and proliferation assay was carried out after 48 h of exposition. Bar charts represent mean, error bars represent SD. * p < 0.0001, ns = not significant, Student’s t test