Fig. 5: POLR2M silences Mir139 in MLL-AF9 cells.
From: The tumor suppressor MIR139 is silenced by POLR2M to promote AML oncogenesis
A Schematic overview of the sgRNA library screen with MLL-AF9 WT (MA9 WT) and MLL-AF9 Mir139KO (MA9 Mir139KO) cells. B Volcano plot showing –log10 P value (Y axis) and average log2 fold change (X axis) of all targeted genes in MLL-AF9 WT relative to MLL-AF9-Mir139KO 14 days post transduction. The red closed circle indicates Polr2m. Red dashed lines indicate a fold change of ±10. Top 10 of enriched genes are shown. C Boxplots showing the reads (n + 1) of six sgRNAs targeting Polr2m in MA9 WT and MA9 Mir139KO cells. The order of the sgRNAs in the MA9 Mir139KO sample corresponds to the same sgRNAs in the MA9 WT sample. Blue circles indicate a loss of ≥10 fold per sgRNA, while red circles indicate a loss of ≤10 fold of the sgRNA. D MLL-AF9 cells and MLL-AF9 Mir139KO cells were transduced with three viruses expressing sgRNAs targeting Polr2m or three viruses expressing control sgRNAs. The left panel shows the colony-forming units (CFU) of Polr2m-targeted MLL-AF9 cells relative to MLL-AF9 Mir139KO cells. The right panel shows the viability of Polr2m-targeted MLL-AF9 cells relative to MLL-AF9 Mir139KO cells (Y axis). The two-tailed unpaired student’s t test with Welch’s correction was used for statistical analysis. The graphs are representative of three independent experiments. E Viability of POLR2M-targeted Molm-13 cells (left panel) or primary MLL-AF9 patient samples (right panel) relative to control targeted cells. The two-tailed paired student’s t test was used for the statistical analysis. Data are representative of two/three experiments. F Expression levels of pri-miR-139 in POLR2M-targeted Molm-13 cells (left panel) or primary MLL-AF9 patient samples (right panel) relative to control targeted cells. The two-tailed unpaired student’s t test with Welch’s correction was used for statistical analysis. The graphs are representative of two/three independent experiments. G Expression levels of pri-miR-139 in POLR2M-targeted U937, MV4-11, TF-1, or HL-60 cells relative to control targeted cells (left panel). Viability of POLR2M-targeted U937, MV4-11, TF-1, or HL-60 cells relative to control targeted cells (right panel). The two-tailed unpaired student’s t test with Welch’s correction was used for statistical analysis. The graphs are representative of two independent experiments. All graphs are depicted as mean ± SEM.
