Fig. 6

DOT1L and NPM1 engage in a feedback loop regulation. A Representative immunoblot analysis of NPM1 expression upon 3 days of NPM1 KD or EPZ treatment (EPZ) in N2a cells. Ponceau staining was used for total protein normalization. B Quantification of the signal intensities shown in A. NPM1 levels were first normalized to the corresponding ACTIN signal and then presented as a fold change over the control values in both conditions (Intensity/CTR). Statistical analysis was performed on n = 9 biological replicates using an unpaired two tailed t-test. *p < 0.05, **p < 0.01. Error bars represent S.D. C Individual data points plot showing the fold change expression (FC OE/CTR) of DOT1L and NPM1 calculated after RT-qPCR upon 3 days of DOT1L OE in N2a cells. Gapdh was used as reference gene for normalization. Statistical analysis was performed on n = 7 biological replicates using a unpaired one tailed (Dot1) and two tailed (Npm1) t-test. *p < 0.05, **p < 0.01. Outliers were identified using a Grubbs method. Error bars represent S.D. D Individual data points plot showing the fold change enrichment of DOT1L (FLAG) over control after by ChIP-qPCR analysis upon 3 days of DOT1L OE in N2a cells. Statistical analysis was performed on n = 3 biological replicates using a paired one tailed t-test: *p < 0.05, **p < 0.01. Error bars represent S.D