Fig. 8

PVT1 knockdown helped to achieve RA amelioration via in vivo experiment. a, the percentage of increase of paw swelling of rats in each transfection group; b, PVT1 expression and mRNA expression of sirt6 in RA-FLSs determined by RT-qPCR; c, protein level of sirt6 in RA-FLSs determined by western blot analysis; d, the gray value analysis of sirt6 in RA-FLSs; e, the serum levels of TNF-α, IL-1β, IL-10 and IL-4 measured by ELISA; f, the mRNA expression of Ki67, PCNA, Bcl-2, Bax and caspase-3 in RA-FLSs assessed by RT-qPCR; g, protein levels of Ki67, PCNA, Bcl-2, Bax and caspase-3 assessed by western blot analysis; h, protein levels of Ki67, PCNA, Bcl-2, Bax and caspase-3 in RA-FLSs; i, the RA-FLS proliferation determined by EDU staining (× 100); j, a bar chart of percentage of EDU positive stained RA-FLSs; k, the RA-FLS apoptosis evaluated by flow cytometry; l, a bar chart of apoptosis rate of RA-FLSs; m, the cell cycle distribution of RA-FLSs assessed by flow cytometry; n, a bar chart of percentage of cell cycle of RA-FLSs; ^* p < 0.05 vs. the control group; ^# p < 0.05 vs. the RA + sh-PVT1-NC group. The results of quantitative analysis were presented as mean ± standard derivation; one-way analysis of variance was performed for comparison; the experimental data was obtained from the average of 3 independent experiments. PVT1, long non-coding RNA plasmacytoma variant translocation 1; 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