Fig. 2.

EZH2 inhibition modulates inflammatory, catabolic, pain-related and autophagic responses in fibroblast-like synoviocytes. Human fibroblast-like synoviocytes were stimulated for 48 h with IL-1β (1 ng/mL). EZH2 mRNA expression was analyzed by RT-qPCR (a). Cells stimulated with IL-1β in the presence or absence of EPZ-6438 (10 µM) were stained for H3K27me3 (FITC, green) by immunocytochemistry, and nuclei were counterstained with DAPI. Representative images from three independent experiments are shown. Quantification of H3K27me3-positive cells is presented (b). The expression of inflammation-, catabolism- and pain-related genes (IL1B, MMP13 and NGF) was analyzed by RT-qPCR (c). Autophagy-associated signal was assessed using a fluorescent autophagosome detection assay (MAK138). Representative images from three independent experiments are shown (d). The MAK138-positive area was quantified and expressed as a percentage of total cell area (e). Data are presented as mean ± SEM from at least three independent biological experiments. Statistical significance is indicated as *p < 0.05, **p < 0.01 and ***p < 0.001