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. 2012 Apr 13;31(10):2350–2364. doi: 10.1038/emboj.2012.98

Figure 2.

Figure 2

EHD2 regulates motility of caveolae. (A) HeLa cells were transfected with siRNA targeting EHD2 or non-targeting siRNA (siRNA control) and cell lysates were analysed after 72 h by SDS–PAGE/western blot using the indicated antibodies. (B) Large regions in the cell periphery of HeLa cells, stably expressing CAV1–EGFP, were bleached and the movement of CAV1–EGFP back into the bleached region monitored through fluorescence recovery. Intensities before and directly after bleaching (t=0) were normalized to 100 and 0%. Cells were imaged every 20 s for 12 min. Recovery curves for cells treated with siRNA control (n=12) or EHD2 siRNA (n=19) are presented. Data points present mean values±s.e.m. (C) Relative CAV1 fluorescence 12 min after bleaching in cells transfected with siRNA control (n=12), EHD2 siRNA oligo1 (n=12) or oligo2 (n=7) or after EHD2 silencing and transfection with EHD2–mCherry rescue (n=10). (D) Relative cavin-1–EGFP signal 12 min after bleaching in HeLa cells, stably expressing cavin-1–EGFP, transfected with siRNA control (n=9) or EHD2 siRNA (n=7). (E) Relative CAV1 fluorescence 8 min after bleaching in HeLa cells transfected with EHD2 siRNA (n=12) or after EHD2 silencing and transfection with dyn-K44A-RFP (n=6). All bars represent mean±s.e.m. Significance of mean differences between the conditions was analysed with a two-tailed unpaired t-test. ***P-values<0.0001.