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. 2011 Jan 14;18(6):959–973. doi: 10.1038/cdd.2010.170

Figure 5.

Figure 5

Mdk promotes resistance to THC-induced autophagy and apoptosis. (a) Effect of THC (2.5 μM, 24 h) on LC3 immunostaining of siC, siMdk or siALK-transfected T98 cells. Values in the upper right corner at each photomicrograph correspond to the percentage of cells with LC3 dots relative to the total number of cells (mean±S.D.; n=3; representative photomicrographs of each condition are shown; **P<0.01 from vehicle-treated, siC-transfected cells; ##P<0.01 from THC-treated, siC-transfected cells). Inset: high-magnification photomicrographs of the arrow-pointed cells. (b) Effect of THC (24 h) on LC3 lipidation of siC, siMdk or siALK-transfected T98 cells. A representative experiment of four is shown. Right panel: densitometric analysis of LC3-II lipidation. Data correspond to LC3-II optical density values (OD) for each condition relative to vehicle-treated, siC-transfected cells (mean±S.D.; n=4; THC 2.5 μM; **P<0.01 from the corresponding vehicle-treated cells; ##P<0.01 from THC-treated, siC-transfected cells). (c) Effect of THC (2.5 μM, 24 h) on apoptosis (as determined by TUNEL) of siC, siALK and siMdk-transfected T98 cells. Data correspond to the percentage of TUNEL-positive cells relative to the total number of cells (mean±S.D.; n=3; **P<0.01 from the corresponding vehicle-treated cells; ##P<0.01 from THC-treated, siC-transfected cells). (d) Effect of THC (1.5 μM, 24 h) on LC3 lipidation of U87 cells incubated with U87 C.M. or T98 C.M. (left panel) or with exogenous Mdk (right panel). A representative experiment of three is shown. Lower panels: data correspond to LC3-II OD for each condition relative to vehicle-treated, U87 C.M.-incubated cells (left panel) or to vehicle-treated cells (right panel; mean±S.D.; n=3; **P<0.01 from vehicle-treated, U87 C.M.-incubated cells (left panel) or vehicle-treated cells (right panel); ##P<0.01 from THC-treated, U87 C.M.-incubated cells (left panel) or THC-treated cells (right panel) and ΦΦP<0.01 from vehicle-treated, T98 C.M. cells (left panel) or vehicle-treated, Mdk-incubated cells (right panel)). (e) Effect of THC (1.5 μM, 24 h) on apoptosis (as determined by TUNEL) of U87 cells incubated with U87 C.M., T98 C.M. or with (+Mdk) or without (−Mdk) exogenous Mdk. Data correspond to the percentage of TUNEL-positive cells relative to the total number of cells (mean±S.D.; n=3; **P<0.01 from the corresponding vehicle-treated cells; ##P<0.01 from THC-treated U87 cells incubated with U87 C.M.- or from THC-treated U87 cells incubated without exogenous Mdk and ΦΦP<0.01 f from vehicle-treated U87 cells incubated with T98-conditioned medium or with exogenous Mdk)