Fig. 1

TRF2 impairs autophagy induced by chemo-therapeutic agents in TNBC. a Human triple negative breast cancer (TNBC) cell lines MDA-MB-231, over-expressing (pTRF2) or not (pBabe) TRF2 were transfected with EGFP-LC3 for 24 hours and treated with Paclitaxel, Carboplatin and Epirubicin for 24 hours at the indicated doses. The autophagic process was evaluated by the quantitative analysis of punctate vesicular structures by fluorescence experiments. Histogram represented the percentage of LC3 puncta positive cells on total GFP positive cells. b Representative images of the experiment described in a were acquired by deconvolution microscopy (63X magnification). Blue: the nuclei stained with DAPI. Green: EGFP-LC3B. c Western blot analysis of LC3 I-II protein levels in MDA-MB-231 pBabe or pTRF2 treated with the indicated drugs in combination or not with 30 µM of the autophagy inhibitor Chloroquine (CQ) for 3 hours. TRF2 protein levels were monitored to check cell lines and b-actin as loading internal control. The histograms represent the mean values ± S.D. of three independent experiments; *p<0.05, **p<0.01, ***p <0.001.