Fig. 6: RSK1 inhibition dampens the monocyte-mediated pro-inflammatory milieu.

a CyTOF analysis of CD14 + monocytes from MF #5 under treatment conditions. Cells were treated with 5 μM ruxolitinib, 5 μM PMD-026, 10 ng/mL TPO, or combination for 4 h. b Changes to cytokine expression from (a) and in four additional MF patients. Comparisons were performed between TPO alone versus TPO + inhibitor. c Immunoblot of CD14 + monocytes from three unique MF patients treated with indicated PMD-026 doses and time points. d Immunoblot of THP-1 cells stimulated with 20 ng/mL TNF and treated with 10 μM PMD-026 alone or in combination for 30 min. Immunoblot is representative of two independent experiments. e Left: Venn diagram showing overlapping DEGs between WashU Cohort MF CD14 + monocytes compared to those from NBM and CMML CD14 + monocytes compared to old healthy controls from (GSE135902). Middle: Key shared candidates from 279 total shared DEGs between MF and CMML monocytes. Right: Top altered Hallmark pathways by enrichment analysis from 279 shared DEGs. Gene count represents the number of genes enriched across each pertinent gene set. Gene ratio represents the number of genes enriched/total number of genes in each gene set. f Immunoblot of CD14 + monocytes from two unique CMML patients treated with indicated PMD-026 doses for 4 h. g qRT-PCR of key effectors from CD14 + monocytes from three CMML patients after 6 h 5 μM PMD- 026 treatment relative to control.