Supplementary Figure 8: Nuclear Mirlet7d reduces cRNA levels by PRC2 mediated transcriptional silencing

a, Top, WB based assay showed specific and efficient EZH2 reduction after transfection of Ezh2-specific shRNA constructs. Bottom, TaqMan based assay showed significant increase of Mirlet7d after transfection of Mirlet7d. b, Top, WB based assay showed specific and efficient EED reduction after transfection of Eed2-specific shRNA constructs. Bottom, TaqMan based assay showed significant increase of Mirlet7d after transfection of Mirlet7d. c, Top, WB based assay showed specific and efficient SUZ12 reduction after transfection of Suz12-specific shRNA constructs. Bottom, TaqMan based assay showed significant increase of Mirlet7d after transfection of Mirlet7d. d, qRT-PCR-based analysis of indicated ncRNA and cRNA in cells transfected with Mirctrl, Mirlet7d or/and Ezh2-specific shRNA constructs. See Fig. 4f-g. e, qRT-PCR-based analysis of indicated ncRNA and cRNA in cells transfected with Mirctrl, Mirlet7d or/and Eed2-specific shRNA constructs. f, qRT-PCR-based analysis of indicated ncRNA and cRNA in cells transfected with Mirctrl, Mirlet7d or/and Suz12-specific shRNA constructs. g, Top, WB based assay showed specific and efficient EZH2 enzymatic activity inhibition after treatment of EZH2-specific inhibitor (EZH2inh, UNC1999). Bottom, TaqMan based assay showed significant increase of Mirlet7d after transfection of Mirlet7d. h, qRT-PCR-based analysis of indicated ncRNA and cRNA in cells transfected with Mirctrl or Mirlet7d or/and treatment with EZH2inh. See also Fig. 4f-g. In a-c, g, representative results from 2 independent experiments (see Supplementary Data Sets 2 and 3). In all plots, data are shown as means ± s.e.m (n=3 independent experiments); asterisks, P values after one–way ANOVA, ***P˂0.001; **P˂0.01; *P˂0.05; ns, non-significant. The statistical test values of each plot are shown in the Supplementary Data Set 4