Fig 2: RhoA overexpression abolishes, while the PTEN inhibitor recovers, the protective effects of CTRP6 against OGD/R-induced inflammation and oxidative stress. Alteration of (A) cell viability and inflammatory cytokines (B) TNF-α, (C) IL-1β, (D) IL-6 and (E) IL-10 levels in different groups (n=3). Alteration of oxidative products (F) ROS, (G) MDA and (H) antioxidant SOD levels in different groups (n=3). **P<0.01 and ***P<0.001 vs. OGD/R + CTRP6 + NC, #P<0.05, ##P<0.01 and ###P<0.001 vs. OGD/R + CTRP6 + RhoA. CTRP6, C1q/tumor necrosis factor-related protein-6; OGD/R, oxygen-glucose deprivation and reperfusion; NC, negative control; RhoA, Ras homolog family member A; PTEN, phosphatase and tensin homologue deleted on chromosome 10; TNF, tumor necrosis factor; IL, interleukin; ROS, reactive oxygen species; MDA, malondialdehyde; SOD, superoxide dismutase.

Fig 3: JC124 treatment diminished TBI-elevated inflammatory cytokine TNF-alpha expression in the brain. Expression level of TNF-alpha in the tissue lysate of ipsilateral cortex (a) and hippocampus (b) was measured with ELISA. In both the ipsilateral cortex (a) and hippocampal samples (b), significantly increased expression of TNF-alpha was only observed in the TBI-vehicle group (p < 0.05 and p < 0.01, respectively). Injured JC124 treated group had significantly lower TNF-alpha expression as compared to vehicle treated group (p < 0.05), and was not different from sham

Fig 4: Siponimod modulates microglial cytokine expression and secretion in an inflammatory milieu. (A,C,E,G) quantitative RT-PCR analysis of Tnf (A), Ifnb (C), Il1b (E) and Il10 (G) gene expression after stimulation for 1 day with +/− 50 µM siponimod +/− LPS. (B,D,F,H) Quantification of TNF-α (B), IFNβ (D), IL-1β (F) and IL-10 (H) protein concentration in the cell culture medium of microglial cells stimulated for 3 days with +/− 50 µM siponimod +/− LPS using respective quantitative sandwich ELISA assays. Grey-dotted lines indicate the lowest standard of the ELISA kit: TNFα = 82.3 pg/mL, IFNβ = 15.63 pg/mL, IL-1β = 68.59 pg/mL and IL-10 = 8.23 pg/mL. Data are presented as mean values ± SEM. Grey dots represent individual data points. Significance of gene expression analysis as well as ELISA was assessed by 1-way analysis of variance (ANOVA) followed by Tukey’s post hoc test using Graph-Pad Prism 8.4.3 (GraphPad Software, San Diego, CA, USA). The experimental groups were considered significantly different at ** p < 0.01, *** p < 0.001.

Fig 5: RT-PCR array with RNA from treated fistulas.(A) Heatmap of a panel of genes relevant to CD. (B) RT-PCR of select genes from the RT-PCR panel confirmed that ADSC-mfNHC-250 inhibited critical proinflammatory cytokine transcripts that participate in the inflammatory cascade, such as TNF-α (TNF-a), IL-6 (IL-6), IL-1α (IL-1a), IFN-γ (IFNg), IL-1β (IL-1b), IL23α (IL23a), Ccr1 (Ccr1), Ccr5 (Ccr5), and Ccl1 (n ≥ 3, *P < 0.05).