Trained monocytes at day 6 were detached using Versene (Gibco), and subsequently washed and counted. Cells were lysed on ice using standard lysis buffer with phosSTOP (Roche, Basel, Switerzland) and cOmplete (Roche), and vortexed every 10 min for half an hour for total lysis, and subsequently stored at −20°C until further use. The frozen homogenate was subsequently centrifuged, and the supernatant was boiled in Laemmli buffer under denaturing and reducing conditions. Trans Turbo Blot System (Bio‐Rad, Hercules, CA, USA) was used according to the manufacturer's instructions. Mini‐PROTEAN Precast Gels (Bio‐Rad) were used, and samples were loaded onto gels together with Precision Plus Protein Dual Color Standard (Bio‐Rad). After running of the gels, proteins were transferred onto nitrocellulose membranes using the semi‐dry method (Bio‐Rad). Next, blots were blocked with 5% milk powder (ELK) in Tris‐buffered saline (TBS) with 0.1% Tween‐20 (TBS‐T; Invitrogen) for 1 h at room temperature. Thereafter, blots were washed in TBS‐T and incubated overnight with an anti‐G9a antibody (Abcam, Cambridge, UK) and anti‐actin (Sigma) on a roller bank at room temperature. Blots were subsequently washed with TBS‐T and incubated for 1h with secondary antibody swine anti‐rabbit‐HRP (Agilent Technologies, Santa Clara, CA, USA), at room temperature. Last, cells were washed and incubated with ECL (GE Health Care, Chicago, IL, USA) before imaging of the blots (ChemiDoc; Bio‐Rad) and analysing (Image Lab version 5.2.1).

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