Fig. 2

NPM1 KD increases H3K79me2 via upregulation of Dot1l. A Representative immunoblot analysis showing an increase in H3K79me2 levels after 3 or 6 days (3D, 6D) of NPM1 KD in N2a cells. Three biological replicates were used (n = 3). B Quantification of immunoblot signal intensities shown in A were conducted using Fiji (ImageJ) software. NPM1 and H3K79me2 levels were normalized to the levels of GAPDH or total H3, respectively. Secondly, the normalized value of each treatment was transformed as a fraction of its control which acquired the value of 1 (CTR = 1) and is shown with a dashed line on the graphs. Error bars represent S.D. Statistical analysis was performed using one sample t-test *p < 0.05, **p < 0.01 (n = 3). C Confocal immunofluorescence showing an example of H3K79me2 staining after 3 days of shNPM1 transfection (NPM1 KD) in N2a cells. (Scale bars 10 µm). D Individual data points plot showing the fold change (NPM1 KD/CTR) expression of the Npm1 and Dot1l genes calculated after RT-qPCR upon 3 days of NPM1 KD in N2a cells. Gapdh was used as reference gene for normalization. Statistical analysis was performed on n = 9 biological replicates using an unpaired two tailed (Dot1l) and one tailed (Npm1) t-test; *p < 0.05, **p < 0.01. Error bars represent S.D