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. 2015 May 12;290(26):16403–16414. doi: 10.1074/jbc.M114.626127

FIGURE 4.

FIGURE 4.

cyt-CRT with various N termini are rapidly degraded by proteasomes. Stably transfected cells expressing CRT-EGFP (A), R-CRT-EGFP (B), or E1V-CRT-EGFP (C) were incubated with MG132 (10 μm) for various durations and analyzed by triple immunofluorescence for detection of EGFP emission using anti-R-CRT pAb and DAPI. Merge images for EGFP emission and anti-R-CRT Ab are shown in the corresponding panels. Scale bars, 20 μm. Quantification of fluorescence intensity in immunofluorescence data of EGFP emission (D) or anti-R-CRT Ab (E) was expressed in relative fluorescence units (RFU). Data are shown as the mean ± S.E. ***, p < 0.001 for intragroup comparison; i.e. gray (or black) bar versus white bar within each of the three experimental groups. ###, p < 0.001 for intergroup comparison with 0 h group; i.e. white, gray, or black bar of 6-h or 12-h group versus the corresponding bar of the 0 h group. N.S., not significant. Results are representative of 180 cells from at least three separate experiments for each condition. F, immunoblotting analysis of CRT-EGFP, R-CRT-EGFP, and E1V-CRT-EGFP content for the indicated MG132 incubation times.