Figure 3
From: Human WRN is an intrinsic inhibitor of progerin, abnormal splicing product of lamin A
Specific duplicated regions in hWRN are critical for senescence. (A) Transfection of GST-tagged WRN-R2 (duplicated peptide) and WRN-R1 (nonduplicated peptide) recombinant proteins into WRN cells (at passage 10) for 24 h. The delivery of WRN-R2 induced H3K9me3 expression in WRN cells compared to WRN-R1 (n = 3 independent experiments; unpaired t-test). (B) The box plots show the intensity of H3K9me3 expression in WRN cells after the delivery of WRN-R1 and WRN-R2. All values are reported as the mean ± SD. (C) Transfection of hWRN and mWRN expression vectors into WRN cells (at passage 10). EV refers to empty vector which was used as a negative control. The expression of Ki67 was induced after transfection with hWRN for 48 h (n = 3 independent experiments; unpaired t-test). (D) Transfection of WRN-R1 and WRN-R2 peptides into HGPS cells (at passage 11) for 24 h. The expression of H3K9me3 was increased in WRN-R2-positive cells (n = 3 independent experiments; unpaired t-test). (E) The box plot shows the intensity of H3K9me3 expression in HGPS cells after the delivery of WRN-R1 and WRN-R2 peptides. Data are the mean ± SD. (F) Transfection of hWRN and mWRN expression vectors into HGPS cells (at passage 11) for 48 h. Transfection of hWRN, but not mWRN, induced H3K9me3 expression in HGPS cells. (n = 3 independent experiments; unpaired t-test). (G) Overexpression of hWRN reduced the expression of progerin and p16INK4A and induced the expression of rad51, cyclin B1, and H3K9me3 in HGPS cells (at passage 11). Blots were cropped from different parts of the same samples and analyzed by film-based imaging systems. **p < 0.001, N.S: not significant. Data are mean ± SD.
