Fig. 2

MCF-7 ALDH1A1 affects in vitro stemness. a Representative images of tumorspheres (4x magnification) showing morphology of spheroids grown on ultra-low attachment plate. Scale bar, 100 μm. b Representative images of tumorspheres (4x magnification) of MCF-7 Scr, MCF-7 ALDH1A1KD and MCF-7 ALDH1A1⁺, showing morphology of spheroids grown on ultra-low attachment plate. Scale bar, 100 μm. b1, b2, b3. Representative images of tumorspheres (10x magnification) of MCF-1 Scr, MCF-7 ALDH1A1KD and MCF-7 ALDH1A1⁺, showing morphology of spheroids grown on ultra-low attachment plate. Scale bar, 100 μm. c Quantification of MCF-7 tumorspheres. Tumorspheres area were calculated using ImageJ Software. Ten pictures for each well were quantified. Tumorspheres> 10.000 pixel square were considered. **p < 0.01 vs MCF-7 Scr. ###p < 0.001 vs MCF-7 ALDH1A1KD. (n = 3). d Western blot analysis of stemness markers CD133 and KLF4 in MCF-7 Scr, MCF-7 ALDH1A1KD, and MCF-7 ALDH1A1⁺ tumorspheres. e Western blot analysis of ALDH1A1, HIF-1α and VEGF in MCF-7 Scr, MCF-7 ALDH1A1KD and MCF-7 ALDH1A1⁺ tumorspheres