Cellular interaction analysis of different KRAB domains with the endogenous TRIM28 protein in human HeLa cells.

24-KRAB fusion constructs, extracts were analyzed by immunoprecipitation (IP) with GST antibodies followed by Western blotting to look for shared complexes with cellular TRIM28 protein. An extract sample (ex) was run as positive control for Western blot immunostaining. The upper part of the blot was probed with anti-TRIM28, the lower part with anti-GST antibodies. A: Example gel for the ZNF10-AB versus A only and ZNF10-PP-AB analysis. B, D: Quantitative evaluation of the blot signals from 3 independent experiments. Columns and numbers indicate relative TRIM28/GST-fusion ratios (ZNF-AB result set to 100%). C: Example gel for the comparison of ZNF10-AB, XFIN-AB and the B subdomain swaps. Statistical significance tested using a one sample 2-tailed T-test. One asterisk p<0.05, two asterisks p<0.01, three asterisks p<0.001.