Skip to main content
. 2015 Oct 7;43(20):9856–9873. doi: 10.1093/nar/gkv1026

Figure 6.

Figure 6.

Overexpression of small RNA binding-deficient Ago2 protein does not Inhibit the nuclear transport of TNRC6A.(A) Cell lysates from HeLa cells transfected with empty vector (pFLAG/HA), pFLAG/HA-Ago2-WT, or pFLAG/HA-Ago2-Y529E were immunoprecipitated with an anti-FLAG antibody. The immunoprecipitates (IP) and cell lysates (input) were analyzed by western blot. (B) MiRNAs contained in the immunoprecipitates of (A) was quantified by real-time PCR. Data are the mean and standard deviation of the triplicate measurements. (C and D) Fluorescent microscopy images of HeLa cells expressing myc-GFP-TNRC6A-NES-mut along with FLAG/HA-Ago2-WT (C) or -Y529E (D). Cells were stained with an anti-HA antibody. Bars, 20 μm. (E) The ratio of cells expressing GFP-signal of myc-GFP-TNRC6A-NES-mut exclusively in the nucleus (N, blue), cytoplasm (C, red), or both (N+C, yellow). (F) Western blot of myc-GFP-TNRC6A-NES-mut and FLAG/HA-Ago2-WT or -Y529E. (G) Cell lysates from HeLa cells expressing myc-GFP-TNRC6A-NES-mut or myc-GFP-TNRC6A-NES-mut-del.GW-I+II along with FLAG/HA, FLAG/HA-Ago2-WT or -Y529E were immunoprecipitated with an anti-FLAG antibody. IPs and inputs were analyzed by western blot.