Etoposide on inducing autophagy of breast cancer cell line MDA-MB-231 and the autophagy process after siRNA knocks down TFDP3 expression.

A, Expression levels of TFDP3 and LC3 in MDA-MB-231 with or without treatment of etoposide for 24 hours using Western Blot; B, Gray scale statistics for fig 4A. The experiment was repeated for three times and the gray scale of Western Blot stripes was analyzed using Image-Pro Plus software; Then an independent sample T-test was applied to analyze the relative gray values between the GFP-LC3 control group and 1μM etoposide treatment group. For the analysis, the relative gray values (TFDP3/β-actin, LC3/β-actin) were calculated; C, Autophagosomes in all groups, and the green fluorescence of GFP-LC3 were evaluated using fluorescence microscopy (n = 3); D, Percentage of the GFP-LC3 positive cells which exhibited punctate GFP-LC3 fluorescence; All groups were evaluated; cells with >2 autophagosomes per cell were declared as exhibiting an autophagic reaction. An independent sample t-test was used to analyze the number of autophagosomes in the control group, which was significantly higher than the siRNA1 group (p<0.05).