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. 2014 Oct 23;10(10):e1004702. doi: 10.1371/journal.pgen.1004702

Figure 5. Mcm4 and Mcm7 transcription and the JH-receptor complex.

Figure 5

(A) Upper panels: western blot (WB) showing the expression of Flag-Met1-3108 and V5-SRC in S2 cells; lower panel: immunoprecipitation (IP) showing the interaction of Flag-Met1-3108 and V5-SRC in the presence of 10 µM JH III or methoprene. α-Flag, Flag antibody; α-V5, V5 antibody. (B) Luciferase assays after co-transfection of pGL4.10/Mcm4 −933 to −37 or pGL4.10/Mcm7 −933 to −11 into S2 cells compared with the pAc5.1 empty vector, pAc5.1/Flag-Met1-3108 (Met) or/and pAc5.1/V5-SRC (SRC), with or without 10 µM JH III or methoprene (JHA) treatment. (C) EMSA using the Mcm4 probe and S2 cell nuclear extracts with expressed Flag-Met and V5-SRC and treated with JH III (10 µM). The arrow indicates the specific complex. FP, free probe. (D) EMSA using the Mcm7 probe and S2 cell nuclear extracts with expressed Flag-Met and V5-SRC and treated with JH III (10 µM). The arrow indicates the specific complex. FP, free probe.