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. 2016 Sep 5;44(20):9803–9820. doi: 10.1093/nar/gkw790

Figure 7.

Figure 7.

AATF(208–560) is functionally active and stabilizes NGDN and NOL10 levels in vivo. (A) An RNAi-rescue experiment was performed in HeLa cells using the localization of ENP1 after LMB treatment (20 nM, 2 h) as readout for nucleolar steps in ribosome synthesis. siRNA treatment was done for 72 h. 24 h after siRNA delivery, cells were transfected with RNAi-resistant constructs encoding for either HA-tagged full-length AATF (fl) or AATF(208-560). Localization of ENP1 and AATF constructs was visualized by IF. Scale bar, 20 μm. (B) Three independent experiments were performed as in (A) and quantified as described in Figure 2C. Mean ± SEM (error bars); n ≥ 58; ***P ≤ 0.001 (unpaired t-test). AATF-HA (fl) and AATF(208-560)-HA significantly complement the AATF depletion effect on nucleolar ribosome biogenesis. (C) RNAi and transfection were performed as in (A), including the AATF(455–560) truncated version. Rescue of NGDN and NOL10 levels was analyzed by IF and nuclei were stained with Hoechst.