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. 2019 Dec 20;27(6):1998–2013. doi: 10.1038/s41418-019-0483-6

Fig. 4. HnRNP F/H knockdown impairs telomerase activity.

Fig. 4

a HeLa cell lysates stably expressing control shRNA (shV) or hnRNP F shRNAs (#1 and #2) were examined for the indicated proteins by WB. b TRAP assays were performed in cell extracts described in a. 500 cells/μl were lysed and 2 μl was used for TRAP assay. IC: Internal Control (36 bp). LB: Lysis Buffer. Error bars represent means ± SD (n = 3). Statistical analysis was performed using Student′s t test (***P < 0.001). c HeLa cell lysates stably expressing control (shV), hnRNP H1 shRNA (#1 and #2), or hnRNP H2 shRNA (#1 and #2) were subjected to WB analysis. d TRAP assays were performed in cell extracts described in c. Error bars represent means ± SD (n = 3). Statistical analysis was performed using Student's t test (**P < 0.01, ***P < 0.001). e HeLa cells stably expressing pLKO.1-vector or pLKO.1-hnRNP F (#1 or #2) co-transfected with siNC or sihnRNP H1/H2 for 72 h, and then transfected with vector or FLAG-tagged DKC1, TCAB1, or hTERT plasmids for 48 h. Then cells were harvested for IP-TRAP assays and WB analysis. WCL, whole cell lysate. f U2OS cell extracts stably expressing control (shV) or hnRNP F shRNA (#1 and #2) were subjected to WB analysis. g U2OS cells described in f were transfected with vector or co-transfected with hTERC and hTERT. 500 cells/μl were lysed and 2 μl was used for TRAP assay. hTERC, hTERT, and GAPDH RNAs were detected by RT-PCR. h HeLa cells described in a transfected with NC (negative control) or hnRNP H1/hnRNP H2 siRNAs were harvested for TRAP assay and WB analysis. Error bars represent means ± SD (n = 3). Statistical analysis was performed using Student's t test (***P < 0.001).