4 male, 4 female LCLs HumanChimpanzeeRhesus Macaque Expression: RNAseq Active Gene Marks: Pol II (ChIPseq) H3K4me3 (ChIPseq) Repressed Region Mark: H3K27me3 (ChIPseq) Enhancer Marks: H3K4me1 (ChIPseq) H3K27ac (ChIPseq) 5 male, 3 female LCLs 6 male, 2 female LCLs 3 Species 8 Samples per species RNAseq+ 5 ChIPseq Figure S1. An illustration of the study design

Figure S2. Choices of cutoffs for classifying regions as enriched

H3K4me1H3K4me3 H3K27acH3K27me3 PolIIRNA Figure S3. Pairwise Spearman’s rank correlations between individuals from the three primates for four histone marks, Pol II and RNA H C R H C R H C R

Figure S4. Fold enrichment of the five marks in regions near TSSs in three primates ChimpanzeeHumanRhesus Macaque ChimpanzeeHuman Rhesus Macaque Fold Enrichment, TSS +/- 10 kb Fold Enrichment, TSS +/- 50 kb A B H3K27ac H3K27me3H3K4me1H3K4me3 Pol II

TSS +/- 2 kb ChimpanzeeHumanRhesus Macaque Figure S5. Pairwise Spearman’s rank correlations between marks in each of the three primates TSS +/- 10 kb TSS +/- 50 kb

Figure S6. Mark enrichment levels are correlated with gene expression levels in chimpanzee R2R2 Scaled Enrichment Level Enrichment Level Low ExpressedMedium ExpressedHigh Expressed Distance to TSS (kb) Gene Expression Level TSS +/- 2kbTSS +/- 10kbTSS +/- 50kb marginal marginal + interactions H3K27ac H3K27me3H3K4me1H3K4me3 Pol II

Figure S7. Mark enrichment levels are correlated with gene expression levels in rhesus macaque R2R2 Scaled Enrichment Level Enrichment Level Low ExpressedMedium ExpressedHigh Expressed Distance to TSS (kb) Gene Expression Level TSS +/- 2kbTSS +/- 10kbTSS +/- 50kb marginal marginal + interactions H3K27ac H3K27me3H3K4me1H3K4me3 Pol II

ChimpanzeeHumanRhesus Macaque TSS +/- 10 kb TSS +/- 50 kb Scaled Enrichment Level Figure S8. Mark enrichment levels are plotted against gene expression levels for sliding windows of genes ordered by increasing expression levels in the three primates Gene Expression Level H3K27ac H3K27me3H3K4me1H3K4me3 Pol II

Predicted Gene Expression Level True Gene Expression Level Figure S9. Scatterplot of predicted gene expression levels against true gene expression levels for all analyzed genes in human H3K27ac H3K27me3H3K4me1H3K4me3 Pol II marginal

H3K4me1 H3K27ac Pol II H3K4me3 H3K27me3H C R Posterior Inclusion Probability Figure S10. Importance of the marginal and first-order interaction effects from the five marks for explaining gene expression levels in the three primates Marginal and Interaction Terms

FDR < 2.5% FDR < 7.5% FDR < 10% Figure S11. TSS regions associated with inter-species differences in enriched marks are enriched for differentially expressed (DE) genes H vs CH vs RC vs R Fold Enrichment of TSS Associated with Inter-Species Mark Differences in DE Genes H3K27ac H3K27me3H3K4me1H3K4me3 Pol II

Figure S12. An example of mark abundance and gene expression levels across three species EENSG , REX02 Human H3K27ac H3K27me3H3K4me1H3K4me3 Pol II RNAInput Rhesus Macaque Chimpanzee

Figure S13. Differences in mark enrichment level plotted against differences in gene expression level for sliding windows of genes ordered by increasing differences in expression level TSS +/- 50 kb Scaled Difference in Enrichment Level Difference in Gene Expression Level H vs R C vs R TSS +/- 10 kb H vs C Scaled Difference in Enrichment Level Difference in Gene Expression Level H3K27ac H3K27me3H3K4me1H3K4me3 Pol II

FDR < 10% FDR < 7.5% FDR < 2.5% Figure S14. R squared by mark enrichment level differences for all pairwise comparisons among the three primates marginal marginal + interactions R2R2 H vs C H vs R C vs R H3K27ac H3K27me3H3K4me1H3K4me3 Pol II

Figure S15. Scatterplot of predicted gene expression level differences plotted against true gene expression level differences for DE genes between human and chimpanzee Predicted Gene Expression Difference True Gene Expression Difference H3K27ac H3K27me3H3K4me1H3K4me3 Pol II marginal

Figure S16. Scatterplot of gene expression level differences plotted against sequence conservation and divergence for DE genes Difference in Gene Expression Level H vs RC vs RH vs C

Figure S17. R squared by four histone modifications and Pol II or by four histone modifications alone H vs CH vs RC vs R w/o Pol II FDR < 5% A C R 2 in Pol II w/ Pol II R 2 in RNA H vs CH vs RC vs R w/o Pol II w/ Pol II HCRHCR R 2 in Pol II R 2 in RNA B D marginal marginal + interactions H3K27ac H3K27me3H3K4me1H3K4me3