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. 2021 Feb 10;13(4):739. doi: 10.3390/cancers13040739

Figure 4.

Figure 4

Effects of MLL2 on H3K4me3 and the mRNA levels of positive regulators of cell cycle. A549 and H1299 cells were transfected with siMLL1, siMLL2, siWDR5, siWDR82, and the off-target control siRNA (siControl). (A) The relative mRNA levels of the targeted genes were measured by qRT-PCR (n = 3, * p < 0.05). (B) The expression level of H3K4me3 was analyzed by western blotting. The uncropped blots of (B) were shown in Figure S4. (C) Levels of H3K4me3 in the siRNAs relative to the siControl. The western blot analysis was performed three times, and the average value was calculated. Error bars indicate the standard deviation (n = 3, * p < 0.05). (D) Cells were transfected with the indicated siRNAs, and the number of cells was calculated using the WST-8 assay on the zero, second, and third days after transfection (n = 4, * p < 0.05). (E,F) The mRNA levels of positive regulatory genes of cell cycle whose H3K4me3 levels were reduced significantly (E) or were not changed (F) in response to metformin were measured by qRT-PCR. Values are expressed as fold changes from siMLL2 to siControl (n = 3, * p < 0.05). The error bars in all bar graphs indicate the standard deviation (n = 3, * p < 0.05).