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. Author manuscript; available in PMC: 2015 Jul 1.
Published in final edited form as: Genes Cells. 2014 May 22;19(7):594–601. doi: 10.1111/gtc.12156

Figure 2.

Figure 2

Expression of wild type and mutant Polκ in normal HF

(A) Western blot analysis of Polκ expression in normal HF cells. Stably expressing 3X Flag-human Polκ was detected by anti-Flag antibody. Actin is used as a loading control. Lane 1, vector control; lane 2, wild type Polκ with siRNA resistant sequence; lane 3, Polκ containing mutations in the PIP1 domain and siRNA resistant sequence; lane 4, Polκ containing mutations in the PIP2 domain and siRNA resistant sequence; lane 5, Polκ containing mutations in the PIP1 and PIP2 domains and siRNA resistant sequence; lane 6, wild type Polκ with siRNA resistant sequence; lane 7, C-terminal deletion of Polκ (1-856) with siRNA resistant sequence. The siRNA target site (siRNA), the polymerase domains, PIP1, PIP2, and UBZ domains are shown in full length (1-870) Polκ. (B) siRNA depletion of 3X Flag-human Polκ in human cells. Cells expressing 3X Flag-wild type or 3X Flag-siRNA resistant mutant Polκ are treated with Polκ siRNA for 48 h. siRNA depletion is determined by Western blotting with anti-Flag antibody. Actin is used as a loading control. siR, siRNA resistant.