Fig. 5

RA-FLS proliferation and inflammation were suppressed and apoptosis was promoted by overexpression of sirt6 in RA. a, the mRNA expression of sirt6 in RA-FLSs determined by RT-qPCR; b, the protein level of sirt6 in RA-FLSs quantified by western blot analysis; c, the protein level of sirt6 in RA-FLSs; d, the serum levels of TNF-α, IL-1β, IL-10 and IL-4 measured by ELISA; e, the mRNA expression of Ki67, PCNA, Bcl-2, Bax and caspase-3 in RA-FLSs evaluated by RT-qPCR; f, the protein levels of Ki67, PCNA, Bcl-2, Bax and caspase-3 assessed by western blot analysis; g, protein levels of Ki67, PCNA, Bcl-2, Bax and caspase-3 in RA-FLSs; h, the RA-FLS proliferation levels determined by EDU staining (× 100); i, EDU positive RA-FLSs assessed by EDU assay; j, the RA-FLS apoptosis evaluated by flow cytometry; k, a bar chart of apoptosis rate of RA-FLSs; l, the cell cycle distribution of RA-FLSs as assessed by flow cytometry; m, a bar chart of cell cycle of RA-FLSs; ^# p < 0.05 vs. the oe-sirt6-NC group; ^& p < 0.05 vs. the sh-sirt6-NC group. The results of quantitative analysis were presented as mean ± standard derivation; one-way analysis of variance was performed for comparison between the treatment group and the control group; the experimental data was obtained from the average of 5 independent experiments. Sirt6, sirtuin 6; RA-FLSs, rheumatoid arthritis fibroblast-like synoviocytes; PCNA, proliferating cell nuclear antigen; Bcl-2, B-cell lymphoma/leukemia 2; Bax, Bcl-2-associated X protein; GAPDH, glyceraldehyde-3-phosphate dehydrogenase; TNF-α, tumor necrosis factor-α; IL-1β, interleukin-1β; IL-10, interleukin-10; IL-4, interleukin-4; EDU, 5-Ethynyl-2′-deoxyuridine; PI, propidium iodide; FITC, fluorescein isothiocyanate; ELISA, enzyme-linked immunosorbent assay; RT-qPCR, reverse transcription quantitative polymerase chain reaction; NC, negative control