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. Author manuscript; available in PMC: 2017 Jun 1.
Published in final edited form as: DNA Repair (Amst). 2016 Apr 21;42:82–93. doi: 10.1016/j.dnarep.2016.04.006

Fig. 1.

Fig. 1

RNF138 interacts with RAD51D. (A) RNF138 clones isolated from yeast two-hybrid screening with RAD51D. (B) Yeast two-hybrid analysis of interactions between RNF138 and RAD51 paralogs. AH109 haploids were co-transformed with the indicated GAL4 DBD and AD fusion constructs and serially diluted on to non-selective and selective growth medium. (C) Protein interaction was quantified by measuring β-galactosidase activity. Data represent mean +/− SEM from three independent experiments conducted in triplicate. (D) Lysates from cells transfected with the indicated expression constructs were subject to anti-HA immunoprecipitation followed by immunoblot analysis with anti-Myc (RNF138, top panel) and anti-HA (RAD51D, bottom panel) antibodies. (E) A reciprocal immunoprecipitation experiment was performed for HA-RNF138 and Myc-RAD51D. (F) Cells were transfected with vectors for either wild-type (WT) or RING finger domain mutant and Myc-RNF138 for anti-HA immunoprecipitation. Abbreviations: DBD Fusion- GAL4 DNA binding domain fusion, AD Fusion- GAL4 activation domain fusion, IP- immunoprecipitate, WC- whole cell lysate, FT- immunoprecipitation flow through, W- immunoprecipitation wash, E- first eluate from immunoprecipitation.