Figure 2

BECN1 translocates to CAV1-located subcellular compartment in response to H₂O₂. (a) HeLa cells were transfected with BECN1-GFP for 24 h, and then treated with 3 mM H₂O₂ for 30 min, EBSS for 2 h and 100 nM rapamycin for 1 h. Cells were fixed with 4% paraformaldehyde for 5 min and observed under a confocal microscope. (b and c) Control (shCtrl.) and CAV1 knockdown (shCAV1) HeLa stable cells were transfected with BECN1-GFP for 24 h. Cells were treated with 3 mM H₂O₂ for 30 min and observed under a confocal microscope (b). The number of cells with BECN1 puncta in confocal images, including (b), was quantified (mean values±S.D., n=10; **P<0.005 versus control cells) (c). (d) HeLa cells were transfected with both CAV1-GFP and BECN1-mRFP for 24 h and then treated with 1 mM H₂O₂ for 30 min. Cells were then observed under a confocal microscope (scale bars=10 μm). (e) The Pearson’s coefficient between CAV1-GFP and BECN1-mRFP determined from confocal images, including (d), was calculated (mean values±S.D., n=5; **P<0.005 versus control cells)