Fig 1: FOXC1 overexpression counteracts the impacts of TRIM22 reduction on the proliferation, apoptosis, migration, invasion and inflammatory response of RA-FLSs. mRNA and protein expression levels of TRIM22 in RA-FLSs transfected with si-TRIM22 and Ov-FOXC1 were detected via (A) RT-qPCR and (B) western blotting. (C) RA-FLS cell proliferation was detected using the Cell Counting Kit-8 assay. (D) Cell apoptosis was assessed using flow cytometry. (E) Western blotting was performed to analyze the protein expression levels of apoptosis-related proteins. The (F) migration and (G) invasion of RA-FLSs were assessed via Transwell assays. Magnification, ×200. (H) Western blotting was performed to analyze the protein expression levels of MMP2 and MMP9. (I) Levels of TNF-a, IL-1ß and IL-6 were assessed using ELISA. (J) RT-qPCR determined the mRNA expression levels of TNF-a, IL-1ß and IL-6. *P<0.05 and***P<0.001 vs. si-NC; #P<0.05, ##P<0.01, ###P<0.001 vs. si-TRIM22 + Ov-NC. FOXC1, Forkhead box C1; TRIM22, tripartite motif-containing 22; RA, rheumatoid arthritis; FLS, fibroblast-like synoviocyte; si, small interfering RNA; Ov, overexpression; RT-qPCR, reverse transcription-quantitative PCR; NC, negative control.
Fig 2: Circ-FBXW12 regulated cell proliferation, inflammation, cell cycle, ECM production and oxidative stress in HG-treated HMCs by targeting miR-31-5p. A The expression of miR-31-5p in HMCs transfected with anti-miR-NC or anti-miR-31-5p was determined by RT-qPCR assay. (B-I) HMCs cells were transfected with si-NC, si-circ-FBXW12, si-circ-FBXW12 + anti-miR-NC or si-circ-FBXW12 + anti-miR-31-5p under HG condition. B The expression of miR-31-5p in HMCs was detected by RT-qPCR assay. C HMC viability was assessed by CCK-8 assay. D The concentrations of IL-6 and TNF-a in HMCs were examined with ELISA kits. E The cell cycle process in HMCs was analyzed by flow cytometry analysis. F The proliferation of HMCs was assessed by EdU assay. G–I The protein levels of CyclinD1, P21, collagen I, collagen IV and TGF-ß1 in HMCs were measured via western blot assay. J and K SOD activity and MDA level in HMCs were detected with specific kits. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001
Fig 3: GNAS knockdown inhibits LPS-induced IL-6 expression by suppressing STAT3 activation in HCC cells. a HepG2 cells were transfected with si-NC or si-GNAS for 24 h and then treated with LPS (5 µg/ml) or not for the indicated hours. Then, the protein expression levels of p65, phosphorylated p65 (p-p65), and GNAS were detected by Western blotting. b HepG2 cells were transfected with pCMV-myc vector or pCMV-myc-GNAS for 24 h, and then treated with the specific inhibitor of NF-kB, PDTC, for 30 min. IL-6 mRNA expression levels were detected by qRT-PCR. c HepG2 cells were transfected with si-NC or si-GNAS for 24 h and then treated with LPS or MED for 30 min. Transcription factors activation profiling plate array was performed. d HepG2 cells were transfected with pCMV-myc vector or pCMV-myc-GNAS for 24 h, and then treated with a specific inhibitor of STAT3, c188–9, for 30 min. IL-6 mRNA expression levels were detected by qRT-PCR. e HepG2 cells were transfected with si-NC or si-GNAS for 24 h and then treated with LPS or MED for 30 min. STAT3 binding on IL-6 promoter was detected by ChIP assay. f HepG2 cells were transfected with si-NC or si-GNAS for 24 h and then treated with LPS (5 µg/ml) or not for the indicated hours. Then, the protein expression levels of STAT3, phosphorylated STAT3 (p-STAT3), and GNAS were detected by Western blotting. Data are represented as means ± SD (n = 3; *represents P < 0.05)
Fig 4: NF-?B agonist reverses the effects of ART on inflammation and oxidative stress of SH-SY5Y cells triggered by OGD/R exposure. SH-SY5Y cells were pre-treated with 100 µM ART, an NF-?B antagonist, or 100 µM ART + NF-?B agonist for 2 h and then exposed to OGD/R. (a–c) ELISA was used to determine the secretion of TNF-a, IL-1ß, and IL-6 in the supernatant of SH-SY5Y cells. (d–g) Activities of SOD, CAT, and GSH-Px and the level of MAD were determined. **p < 0.01 vs OGD/R; ## p < 0.05, 0.01 vs OGD/R + ART.
Fig 5: LIN28A aggravated HG-induced injury by targeting Nox4. HK-2 cells were transfected with si-NC, si-LIN28A, si-LIN28A + vector or si-LIN28A + oe-Nox4 and subsequently treated with HG for 48 h (A) Western blotting analysis of LIN28A and Nox4 (n=4). (B) qRT-PCR analysis of Nox4 (n=4). (C) Cell viability was examined by MTT assays (n=4). (D) TUNEL staining (TUNEL, green. Scale bar=100 µM) were examined. (E) Cellular ROS levels were examined by a fluorometric ROS sensor (red, n=4. Scale bar=100 µM). (F) Secretion of TNF-a and IL-6 into culture supernatants (n=4). *P < 0.05, **P < 0.01 and ***P < 0.001.
Supplier Page from Abcam for Human IL-6 ELISA Kit