Fig 2: Effect of Djulis hull crude extract on the protein expression of (A) PPAR?, IL-1ß, IL-6, and TNF-a, (B) SIRT1 and LXRa, and (C) lipogenesis and ß-oxidation pathways in the livers of HFD-induced obese mice. ND: normal diet; HFD: high-fat diet; HCE: high dosage of crude extract. Values represent the mean ± SEM (n = 6). Statistical methods were used by one-way ANOVA, and the different letters are significantly different at p < 0.05.

Fig 3: STING inhibition reduced inflammatory and degenerative phenotypes of NP cells induced by H2O2. (a–d) NP cells were pre-exposed to negative control siRNA or siSTING followed by 500 μM H2O2 treatment for 48 h. The relative mRNA amount of TNF-α (a), IL-1β (b), IL-6 (c) was determined by qPCR (n = 4 independent experiments). (d) Representative Western blot images of collagen II, aggrecan, Mmp3, Mmp13 protein expression in treated NP cells from three independent experiments. (e–g) NP cells were pre-exposed to negative control siRNA or siSTING followed by 500 μM H2O2 treatment for 2 h. (e) Representative Western blot images of STING–NF–κB pathway expression in treated NP cells from three independent experiments. (f) Representative IF staining of p65 in treated NP cells. (g) Quantitative analysis of cells with nuclear p65 in IF images (n = 4 independent experiments). The values presented are the means ± SDs. *P < 0.05; **P < 0.01; ***P < 0.001; ns, not statistically significant. Statistical significance was assessed by one-way ANOVA with a post-hoc Tukey test.

Fig 4: The role of Ang1 in IL-6 mediated Bev anti-angiogenic effect enhancement. (A) Images of tubes formed by HUVEC in the conditioned media from RMG-1 treated by indicated siRNA and anti-IL-6 antibody (magnification × 40). Fluorescent microscope observation was made after 18 h of incubation and Calcein AM staining. (B) Tubes in each well with indicated condition were measured by hybrid cell count software in order to elucidate the tube area. Data were average of triplicated well. (C) Reduction rate of tube area by Bev treatment with or without IL-6 blockade and siRNA, IL-6 blockade weakened Bev function in siCTL whereas siAng1 restored the anti-angiogenic function. Data are shown from one of two independent experiments with similar results. siCTL: siRNA without gene silencing ability (control). siAng1: angiopoietin-1 silencing siRNA. Error bars are SEs. All the images were observed by fluorescence microscope (BZ-X800, Keyence, Osaka, JAPAN). Then, these obtained images were analyzed by the BZ-H4C analytic application (Keyence) for hybrid cell count and the BZ-H4CM application (Keyence) for macro cell count.

Fig 5: Effects of Epi on LPS-induced inflammatory cytokine production. (a) Chemical structure of (-)-Epicatechin. (b) Effects of different concentrations of Epi alone on cell viability. (c) Experimental scheme: BV2 microglial cells were pretreated with DMSO or Epi for 0.5 h, and then cells were treated with normal saline or LPS for 12 h. (d) Expression of IL-6, IL-1ß, and TNFa at the mRNA level in cells 12 h after LPS induction (n = 3 for each group). (e–h) Determination of IL-6, IL-1ß, and TNFa protein levels by western blot analysis. n = 3 for each group. All data are shown as mean ± SEM. One-way ANOVA test followed by Tukey's post hoc test, *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.