Supplemental Figure 1 Figure S1. KDM4A activity: enzymatic evaluation. Schematic representation of KDM4A peptide-based fluorescent screening chemistry. First reaction with H3K9me2 and formaldehyde production (i); second step of the reaction, where the formaldehyde generated in (i) reacts with ammonia and 2 molecules of acetoacetanilide (ii).

Figure S2 Additional cellular effect of PKF118-310 Figure S2 Additional cellular effect of PKF118-310. (A) Doubling time rate of HCT-116 wild-type cell line (dark blue), cells treated with PKF118-310 at 10 μM (light blue) and 1 μM (purple), background (red) and cells without FBS (green). (B) Duplicate proliferation curve relative to HCT-116 cell line with the color scheme as reported above. Supplemental Figure 2

A B Figure S3. KDM4A activity: cellular evaluation. (A) Cell cycle analysis of PKF118-310 from 0.01 μM to 10 μM for 24h in HCT-116 p53-/- cells. (B) Western blot of U937 histone extract incubated with H3K9me1, H3K9me2 and H3K9me3, respectively. (*) indicates the marker position. Supplemental Figure 3

HCT-116 Figure S4. H3S10ph induction mediated by PKF118-310. Western blot of HCT-116wt histone extract incubated with H3S10ph. H3S10ph H4 DMSO 12h 24h 48h 24+24h Supplemental Figure 4