Fig. 4

TGF-β1-induced EMT represses DOT1L in H358 cells. Cells were treated with TGF-β1 (10 ng/ml) for 48 h. a DOT1L mRNA expression was measured by RT-qPCR and normalized to GAPDH mRNA. The graph corresponds to the mean ± SD of three independent experiments with PCR in duplicate. **p = 0.0056 by Student’s t test. b DOT1L protein was analyzed by immunoblotting of A549 and H358 total cell lysates. Actin was used as a loading control. The apparent molecular weights (kDa) are indicated on the right of the panel. This blot is representative of four independent experiments. c The graph corresponds to the quantification of the intensity of the protein bands for A549 and H358 cell lines with (white bars) or without TGF-β1 (black bars) treatment after normalization to actin for DOT1L and total histone H3 for H3K79me3. Value 1 corresponds to untreated cells. Statistical analysis was performed for four independent experiments with the Student’s t test (** corresponds to p < 0.01)