Figure 4

Ectopic expression of flotillins normalizes EGFR expression in stable MCF7 flotillin knockdown cells. MCF7 control and flotillin-1 (sh-F1-A/B) or flotillin-2 (sh-F2-B) knockdown cells stably transfected with flotillin-2 (Flot2-EGFP) and flotillin-1 (Flot1-resA/-resB) constructs were grown subconfluent on coverslips, fixed and stained with antibodies against endogenous EGFR (red) and GFP (green). EGFP was stained with antibodies to enhance the signal as the expression levels in the stable cells were kept low in order not to exceed that of endogenous flotillins in normal cells. Nuclei were stained with DAPI. Scale bar: 10 μm.