Fig. 2.

Digitoxin and MonoD treatment modulate levels of autophagy markers and autophagy regulatory proteins. A: H460 cells were treated with 50 nM Digitoxin and MonoD for 1 h, lysed and 40 μg lysate was separated on a 10% SDS-PAGE and assayed for autophagic marker LC3-II levels by immunoblot analysis. Blots were re-probed with GAPDH antibody to confirm equal loading of the samples. B: Densitometric quantification of LC3-II/LC3-I levels in response to drug treatment. C: H460 cells treated with 50 nM Digitoxin and MonoD for 1 h were lysed and assayed for autophagy regulatory proteins such as Beclin-1, NF-κB and p38 MAPK levels by immunoblot analysis. Blots were re-probed with GAPDH antibody to confirm equal loading of the samples. D: Beclin-1, NF-κB and p38 MAPK blots were quantified with densitometry using ImageJ. (*P < 0.05 for each drug treated data point as compared to non-treated control). E: H460 cells were treated with 50 nM Digitoxin and MonoD for 1 and 6 h, lysed and 40 μg lysate was separated on a 10% SDS–PAGE and assayed for autophagic marker Beclin-1, p38 MAPK, and NFkB levels by immunoblot analysis to demonstrate the temporal change in autophagy regulatory proteins. Blots were re-probed with GAPDH antibody to confirm equal loading of the samples.