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. 2012 Jul 17;122(8):2827–2836. doi: 10.1172/JCI62374

Figure 1. PHD3 interacts with HCLK2.

Figure 1

(A) Flag-PHD3 immunoprecipitated from HeLa cells was separated by SDS-PAGE and stained with Coomassie blue. Protein identities were confirmed with MALDI/TOF/TOF. (B) HeLa cells transfected with Flag–PHD1–PHD3 or empty vector were harvested after treatment with DMOG or PBS for 4 hours. Immunoprecipitation with anti-Flag beads was followed by Western blot analysis with the indicated antibodies. (C) Truncated Flag-PHD3s or (D) truncated Flag-HCLK2s and GFP-PHD3 were transfected into HeLa cells. Cell lysates were immunoprecipitated with anti-Flag beads, and Western blot analysis was performed with the indicated antibodies. (E) GST or GST-HCLK2340–530 fusion proteins (1 μg) were incubated with an equal amount of recombinant PHD3 prior to GST pull down. Pull-down product and 5% of PHD3 input were separated with SDS-PAGE and analyzed by Western blot with anti-PHD3 or anti-GST antibodies. (F) Endogenous PHD3 was immunoprecipitated with an anti-PHD3 antibody. Immunoprecipitates were analyzed by Western blot with anti-PHD3 or HCLK2 antibodies. Representative blots from at least 3 experiments are shown in B–D.