G9a Inhibition Induces Autophagic Cell Death via AMPK/mTOR Pathway in Bladder Transitional Cell Carcinoma
Fig 4
G9a inhibition induces autophagic vacuoles in T24 and UMUC-3.
(A and B) Morphological changes of T24 and UMUC-3 cells after treatment of 1.5 μM BIX-01294 for 24 h or UMUC-3 cells transfected by shG9a #1 for 72 h by inverted phase contrast microscopy (×100 and ×200). Arrows point to cytoplasmic vacuole accumulation. (C) Representative electron micrographs of T24 cells treated with 1 μM BIX-01294 for 24 h and 72 h transfected UMUC-3 cells (×8000 and ×40,000). Arrows point to distinct autophagic structures. (D) Examples of cells transiently transfected with ptfLC-3 plasmid and treated with 1 μM BIX-01294 for 24 h or transfected with shG9a #1 for 72h under fluorescence microscope (×200). Yellow arrows point to autophagosomes while red arrows point to autolysosomes. (E) Quantification of the number of autophagosomes (yellow LC-3 puncta) and autolysosomes (red LC-3 puncta) per cell. Rectangle indicates the magnifying picture right to it in the same treatment group. *P < 0.01.