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. 2015 Feb 9;16(2):3705–3721. doi: 10.3390/ijms16023705

Figure 4.

Figure 4

hDrn1 localizes to the nucleoplasm in HeLa cells by immunofluorescence and co-immunoprecipitates with endogenous hDbr1. (A) The expression vector encoding myc-tagged hDrn1 was transfected into HeLa cells. Twenty-four hours after transfection, the cells were stained with anti-myc (M192-3, MBL, panel FITC, middle), anti-hDbr1 16019-1-AP, Proteintech) and Hoechst 33342 (SIGMA, panel Hoechst, bottom) to label the nuclei. Differential interference contrast (DIC) image of the cells is also shown as Phase panel at the top; and (B) Co-immunoprecipitation of endogenous hDbr1 with Flag-hDrn1 in vivo. Flag-tagged hDrn1 was expressed in HEK293T cells and immunoprecipitated using anti-Flag M2 antibody (SIGMA). Co-precipitated fractions were separated using 12.5% SDS-PAGE gel and analyzed by Western blotting using anti-hDbr1 and anti-Flag polyclonal antibodies (upper panel) and anti-β-actin antibody (MBL) (lower panel). Input lanes contain 5% of the total proteins used for immunoprecipitation assays. The positions of protein mass markers are shown on the left in kDa.