Fig 2: Set7 Regulates Metabolic Changes in Cells Trained with ß-Glucanß-Glucan stimulation activates Set7 to write the H3K4me1 modification to distal enhancers associated with sustained MDH2 and SDHB gene expression leading to increases in TCA cycle metabolites and increased oxidative phosphorylation important for enhanced cytokine production in trained immunity.

Fig 3: Set7/9 and PDX-1 promoted the effects of ISL-1 on ß-cell proliferation. (A, B) The proliferation of NIT-1 cells transfected with the indicated plasmids in 96-well plates was analyzed with CCK-8 kits. (C, D) NIT-1 cells in 6-wells plates were transfected with the indicated plasmids. The cell confluence was measured using the Genetix clone selector after 48 h. The data represent 3 independent experiments, each performed in triplicate. *p < 0.05, **p < 0.01 vs. pcDNA3, #p < 0.05, ##p < 0.01 vs. non-silencer, NS, non-significance. +p < 0.05, ++p < 0.01 as indicated.

Fig 4: Characterization of constitutive KD of SetD7 in INS-1 cells and temporal measurements of SetD7 expression in a mouse model of HFD-induced metabolic stress.A, low power overview of live INS-1 cells expressing RFP-tagged SetD7 shRNA or scrambled (Scr) shRNA showing RFP positive expression of scrambled and shRNA against SetD7 expression. B, representative WB and band quantitation for SetD7 and Pdx1 in SetD7 KD INS-1 cells relative to the scrambled control, *p < 0.01. C, measurement of the glucose-stimulated insulin secretion (GSIS) in the scr control versus SetD7 KD INS-1 cells. The INS-1 cells were primed for GSIS following 12 h incubation in the complete RPMI media with 2.8 mM glucose followed by equilibration for 2 h in warm and oxygenated KRBH pH 7.4. The GSIS was performed with low glucose (2.8 mM) and high glucose (16.7 mM) for 1 h in KRBH pH 7.4. Secreted insulin was measured by ELISA. The secreted insulin data was normalized with total protein content in each well. The scr shRNA control INS-1 cells under high glucose (16.7 mM) exhibited an ~4.8-fold increase in GSIS (*p < 0.002), whereas SetD7 KD INS-1 cells demonstrated a modest ~2.1-fold increase in GSIS (**p < 0.001). D, proliferation frequency assay in scr control versus SetD7 KD INS-1 cells measured by BrdU incorporation, *p < 0.0001. E, SetD7 mRNA levels were evaluated (1–16 weeks) in isolated islets of mice undergoing control diet or high-fat feeding. Total RNA preparations were processed for RT-PCR for the expression of SetD7 using FAM-tagged rat PCR primers. The CT values were normalized with ß-actin. The SetD7 mRNA expression data presented is relative to age-matched control chow-fed mice and demonstrates a peak in islet SetD7 at 8 weeks HFD corresponding with the height of ß-cell functional compensation followed by a marked reduction by 16 weeks corresponding with ß-cell decompensation (35). #p < 0.0001 compared with all other time points; ##p < 0.05 compared with weeks 3–8.

Fig 5: Silencing of SET7/9 leads to changes in the expression of E2F1 downstream targets. Downregulation of E2F1, cyclin E1, cyclin A2, and CDK2 was observed in both the SET7/9-silenced and the MTA-treated cells.