Figure 11. JCTH-4 induces autophagy in OS cells alone and with CC.

The presence of autophagic vacuoles in (A) Saos-2 after 96 hours of treatment and (B) U-2 OS cells after 72 hours of treatment with JCTH-4, CC, and solvent control (Me₂SO) was determined by MDC staining. Bright blue punctate marks are indicative of autophagic vacuoles. Corresponding phase and PI micrographs are shown below the MDC images. Scale bar = 15 µm. All images are representative of 3 independent experiments. (C) Cell lysates of Saos-2 cells treated with JCTH-4, CC, TAM as positive control, and solvent control (Me₂SO) for 72 hours were examined for the conversion of LC3-I to LC3-II by western blot analyses. β-actin was probed to serve as a loading control. Densitometric analyses were done using ImageJ software and statistics were calculated using GraphPad Prism version 5.0. Image is representative of 3 independent experiments demonstrating similar trends. Values are expressed as mean ± SD of triplicates of one independent experiment. *p<0.05, **p<0.01 versus solvent control (Me₂SO); #p<0.01 versus 5 µM CC.