Figure 2: Tctp binding to Brm negatively modulates Brm ATPase activity. | Nature Communications

Figure 2: Tctp binding to Brm negatively modulates Brm ATPase activity.

From: Antagonistic roles of Drosophila Tctp and Brahma in chromatin remodelling and stabilizing repeated sequences

Figure 2

(a) Enhanced ARP4/Act binding to Brm by Tctp reduction. V5-tagged ARP4 and Act were transiently expressed in S2 cells, stably transfected to produce 2XFLAG-BrmWT. Tctp-knockdown by RNA interference (RNAi) using double-stranded RNA (dsRNA) was treated twice at day 0 and 3 to maximize RNAi effect. Immunoprecipitation was performed using FLAG antibody. ARP4/Act binding to Brm was increased by Tctp reduction (arrows). (b–d) in vitro ATPase activity assay in Tctp-knockdowned S2 cells. S2 cells were stably transfected to express 2xFLAG-BrmWT. (b) dsRNA was treated twice at day 0 and day3. (c–d) Brm immunoprecipitants from nuclear extracts with low levels of Tctp (arrowheads) were monitored for ATPase assay, relative to control precipitant (pSK(−) RNAi). (d) Brm ATPase activity was increased up to about 2.5-fold by Tctp-knockdown, compared with pSK(−) RNAi control. ATPase activity was normalized to that of pSK(−) control. n=3 tests (mean±s.d.).

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