Fig 2: Effects of TRF365 binding to the 3'-UTR of IKBKB on translation.A qRT-PCR and (B) western blotting evaluation of the effects of co-transfection of an IKBKB siRNA and a TRF365 inhibitor. C Evaluation of the cell apoptosis rate by flow cytometry after 48 h. D Schematic illustration showing the flowchart of the prediction of the TRF365-binding site in IKBKB using a mutant. E Relative luciferase activity. *P < 0.05, **P < 0.01, ***P < 0.001.

Fig 3: Expression of IKBKB in OA and NA ACL cells.A, B IKBKB expression detected by immunofluorescence. C, D Expression of IKBKB in OA and NA ACL cells, with and without IL-1β stimulation. *P < 0.05, **P < 0.01, ***P < 0.001.

Fig 4: Effect of emodin on the expression of inflammatory factors mRNA in the brain tissue of MCAO model rats. (a–d) represent the mRNA expression of IL-6, IL-1β, TNF-a, and TGF-β1,respectively. (e–h) NF-κB and IKKB protein electrophoresis and statistical histogram. Data are presented as mean ± SEM; *P < 0.05, **P < 0.01 vs sham group; #P < 0.05, ##P < 0.01 vs model group.

Fig 5: Human lung ADCs lacking CHUK/IKKa are highly correlated with elevated TNF/TNFRSF1B-associated FOXP3 Treg-cell and macrophage scores. (A, Left) The array containing 90 human lung ADCs (BCS04017a; US Biomax, Inc.) was divided into two groups of low CD68 (L-CD68)- and high CD68 (H-CD68)-positive cells, analyzed by immunofluorescence (IF) staining. (Right) The correlation between macrophages (CD68) and Treg-cell (FOXP3) numbers was further analyzed. n, case numbers; ****P < 0.0001; Student’s t test. (B) Correlation of IKKa expression and macrophage numbers in the human lung ADC array (BCS04017a) was analyzed by IHC staining. The distribution of IKKa-stained intensities from strong (+++) to negative (-) staining was divided into two groups of L(low)-CD68 and H(high)-CD68. Data represent mean ± SEM. Data were analyzed by ?2 test. n, ADC numbers; ***P < 0.001. (C) Comparison of CD68 and CD163 expression patterns in human lung ADC and its adjacent tissue in human tissue arrays (HLung030PG02; US Biomax, Inc.), analyzed by IHC staining. Dark brown, positive cells; numbers on the Top, positions in the tissue array. (Scale bar, 50 µm.) (D) Correlation of CHUK expression levels with macrophage (Left) and Treg-cell (Right) scores in human lung ADCs determined by analyzing the data (14). Using quartile analysis, CHUK expression was divided into high (H-CHUK) and low (L-CHUK) expression levels. CHUK, IKKa; n, ADC numbers per group; P value at the Top of panels, Student’s t test. (E) Correlation of IKBKB expression levels with macrophage (Top) or Treg-cell (Bottom) scores in human lung ADCs determined by analyzing the data (14). Using quartiles and median analyses, IKBKB expression was divided into high IKBKB (H-IKBKB) and low IKBKB (L-IKBKB) expression levels. n, ADC numbers; P value at the Top of panels, Student’s t test. (F) Coexpression of the genes encoding molecules for macrophage and Treg-cell development in human lung ADCs determined by analyzing the data (6). P value at the Top of panels, two-tailed t test; n, ADC numbers; red lines, repression lines; r, Pearson correlation coefficient; -4 (-2 or -1), outliers that were removed from 230. (G) Correlation of H-CHUK or L-CHUK expression levels (using median and quartile analysis to divide CHUK expression groups) with the expression of CSF1, CSF1R, FOXP3, TNFRSF1B, TNF, or CCL22 genes in human lung ADCs (TCGA, PanCancer Atlas, cBioPortal). n, ADC numbers. P value at the Top of panels, Student’s t test.