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. 2022 Dec 2;13:7444. doi: 10.1038/s41467-022-35070-2

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© The Author(s) 2022

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 3 — a Pie chart representing the number of genes down- (log₂FC ≤ −0.5; FDR ≤ 0.05) and up-regulated (log₂FC ≥ 0.5; FDR ≤ 0.05) in Dot1L cKO CMs at E16.5. b Quartile distribution of gene expression in CMs at E16.5. Genes downregulated (Down) in Dot1L cKO were expressed at a high level in control CMs, whereas the majority of upregulated genes (Up) belonged to the bottom quartile of expression. Genes not significantly modulated (Unch) were evenly distributed across quartiles of expression. Data are shown as stacked percentage bar graph. c Heatmap showing the expression of the top 25 transcription regulators downregulated in Dot1L cKO CMs at E16.5, highlighting that multiple chamber-specific transcription regulators were significantly downregulated. RNA-scope analyses (d) and respective quantification (e) validating blunted expression of Hand1 in Dot1L cKOs both at E10.5 and E16.5. (N = 3 biological replicates; unpaired t-test, two-sided, E10.5 RV P = 0.0042, E10.5 LV P = 0.0020, E16.5 LV P = 0.0003). f Quantification of RNA-scope analysis validating no changes in expression of Hand2 in Dot1L cKOs both at E10.5 and E16.5 (RNA-scope images presented in Supplementary Fig. 4) (N = 3 biological replicates). RNA-scope analyses (g) and respective quantification (h) validating reduced levels of Irx4 in Dot1L cKOs both at E10.5 and E16.5. (N = 3 biological replicates; unpaired t-test, two-sided, E10.5 LV P = 0.0153, E16.5 RV P = 0.0006, E16.5 LV P = 0.0005). i Quantification of RNA-scope analysis validating reduced levels of Smyd1 in Dot1L cKOs both at E10.5 and E16.5 (RNA-scope images presented in Supplementary Fig. 3). (N = 3 biological replicates; unpaired t-test, two-sided, E16.5 RV P = 0.0057, E16.5 LV P = 0.008). For panels d and g scale bars = 250 μm for low magnification panels and 50 μm for high magnification images. In panels e, f, h and i data is presented as mean ± SEM; * represents P ≤ 0.05, **P ≤ 0.01. (N = 3 biological replicates). Source data are provided as a Source Data file.