1. Characterising the CMTR1–DHX15 interaction.
Characterising the CMTR1–DHX15 interaction. (A) HeLa cells were transfected with two independent CMTR1 or DHX15 siRNAs or non-targeting control. After 2 d cell extracts were analysed by Western blot. The anti-CMTR1 antibody recognised a 110-kD band, the molecular weight of CMTR1, which was reduced following transfection of CMTR1 siRNAs. (B) CMTR1 was immunoprecipitated from HeLa cell extracts with indicated amount of anti-CMTR1 antibody. Western blot analysis was performed on supernatant or IPs. (C) CMTR1 was immunoprecipitated from Hela cell extracts. Sheep IgG was used for control antibody; Western blot analysis. (D) DHX15 was immunoprecipitated from Hela cell extracts; Western blot analysis. (E) 100 ng recombinant His6-DHX15 and/or CMTR1 were resolved by SDS–PAGE and Coomassie Blue-stained. (F) HeLa cells were transfected with pcDNA5 HA-CMTR1, GFP-CMTR1 or GFP (G). Anti–GFP-antibody IPs were performed. Western blot analysis. (G) As in (F) except pcDNA5 HA-DHX15 and GFP-DHX15 were transfected. (H) 2 μg HeLa cell RNA incubated with or without RNaseI and A and resolved by electrophoresis. HC, heavy chain; LC, light chain.
Francisco Inesta-Vaquera et al. LSA 2018;1:e
© 2018 Inesta-Vaquera et al.