Fig. 6

Analysis of TET activity. a Western blot for TET enzymes (donor Hm14) showing a decrease in TET2 protein levels during the first day of differentiation. Multiple isoforms are detectable for TET2. TET3 levels did not change. TET1 was undetectable by western blotting using commercial antibodies. b TET activity during ​monocyte-to-macrophage differentiation. TET activity in nuclear extract decreases significantly (p < 0.05, ANOVA) during differentiation from freshly isolated monocytes at 0 h to adherent phagocytic cells at later time points (n = 3–5, mean ± SEM). c Quantification of DMR33 methylation after 1-day differentiation exposed to different concentrations of the TET inhibitor Octyl-2-α hydroxyglutarate (2-HG). Results are mean ± SD from 3 independent donor samples (Hm15, HU2 and HU3). **p value <0.01 (unpaired Student’s t test). (D) Light microscopy of monocytes after 1 day differentiation in the presence or absence of TET inhibitor (donor Hm15). While control cells acquire a macrophage-like, adherent phenotype with protrusions, cells treated with 630 µM 2-HG are only loosely attached and have a round phenotype