Fig. 4.

Representative TEM photomicrographs (a) and corresponding fluorescence microscopic images (b) portraying the autophagosomes accumulation in GCNF (25 μg/ml) exposed cells. In TEM image, inset of the red marked area present detailed structure of autophagosomes whereas in fluorescence images yellow arrows marked the GFP-LC3 puncta. Scale Bar – 20 μm. Per view 5 cells and 10 views per group were analyzed. (c) Quantification of autophagosomes per cell in fluorescence images. Values are expressed as mean ± SE of three independent experiment. *p<0.05 was considered as statistical significant. (d –1, 2) Effect of GCNF on autophagic proteins in dose dependent and time dependent manner analyzed through western blot. GAPDH was served as a loading control. Densitometry analysis is represented by the digits given above the respective blots and also provided in Additional file 2: Figure S2a, b. Values are expressed as mean ± SE of three independent experiment. *p<0.05 was considered as statistical significant. (e) Monodansyl cadaverine (MDC) staining of GCNF exposed cell. Scale Bar - 50 μm