Journal: Cell Communication and Signaling : CCS

Article Title: The pathogenic role of succinate-SUCNR1: a critical function that induces renal fibrosis via M2 macrophage

doi: 10.1186/s12964-024-01481-5

Figure Lengend Snippet: Succinate stimulated macrophage infiltration, activation of profibrotic M2 phenotype, upregulation of profibrotic factors in vivo and in vitro. A F4/80 immunohistochemistry staining indicated succinate markedly increased renal macrophage in the kidney interstitium rather than glomerulus. *** P < 0.001, versus control group, n = 5. B Renal proinflammatory M1 cytokines, including iNOS and IL6 mRNA levels, were reduced by succinate. ** P < 0.01, versus the control group, n = 5. C The mRNA levels of anti-inflammatory M2 cytokines (Arg1, Fizz1, Mgl2, and IL-10) in the kidney were markedly increased. ** P < 0.01;*** P < 0.001, versus control group, n = 5. D Succinate upregulated renal M2 macrophages-related profibrotic factors expression (galectin3, MMP9, MMP12, MMP13, PDGF, and CTGF). ns, not significant; ** P < 0.01;*** P < 0.001, versus control group, n = 5. RAW 264.7 cells were treated at 500 μM succinate for 24 h, and quantitative PCR analysis and immunoblotting were adopted to detect the effects of succinate on M2 polarization and expression of profibrotic factors in vitro. E Succinate had no effects on the cell viability of RAW 264.7. Succinate downregulated M1 cytokines (iNOS and IL6) mRNA levels ( F ) while significantly upregulated M2 cytokines (Arg1, Fizz1, Mgl2, and IL-10) ( G ). *** P < 0.001, versus control group, n = 3, biologically repeated 3 times. Likewise, succinate remarkably increased M2 macrophage-related profibrotic factor expression (MMP9, MMP12, MMP13, PDGF, and CTGF) ( H ). ns, not significant; *** P < 0.001, versus control group, n = 3, biologically repeated 3 times. The cell supernatant was measured by mouse CTGF ELISA kit according to the manufacturer’s instructions (E-EL-M0340; Elabscience, Bethesda, MD). Similarly, succinate increased CTGF release of macrophages ( I )

Article Snippet: The cell supernatant was measured by mouse CTGF ELISA kit according to the manufacturer’s instructions (E-EL-M0340; Elabscience, Bethesda, MD).

Techniques: Activation Assay, In Vivo, In Vitro, Immunohistochemistry, Staining, Control, Expressing, Real-time Polymerase Chain Reaction, Western Blot, Enzyme-linked Immunosorbent Assay

Journal: Cell Communication and Signaling : CCS

Article Title: The pathogenic role of succinate-SUCNR1: a critical function that induces renal fibrosis via M2 macrophage

doi: 10.1186/s12964-024-01481-5

Figure Lengend Snippet: CTGF neutralizing antibody inhibited the stimulation of fibroblasts by macrophage-conditioned medium. A CTGF antibody prevented the proliferative effects of CM on NRK-49F, indicated by the results of the CCK8 assay. *** P < 0.001, versus control group, n = 6 in CCK8, biologically repeated 3 times. B Also, the CTGF antibody suppressed the activation effects of CM on NRK-49F, as indicated by the results of the protein quantitative analysis of fibronectin and α-SMA. *** P < 0.001, versus control group, &&& P < 0.001, versus the succinate group, n = 3, biologically repeated 3 times

Article Snippet: The cell supernatant was measured by mouse CTGF ELISA kit according to the manufacturer’s instructions (E-EL-M0340; Elabscience, Bethesda, MD).

Techniques: CCK-8 Assay, Control, Activation Assay

Journal: Cell Communication and Signaling : CCS

Article Title: The pathogenic role of succinate-SUCNR1: a critical function that induces renal fibrosis via M2 macrophage

doi: 10.1186/s12964-024-01481-5

Figure Lengend Snippet: Succinate promoted CTGF expression through activation of β-catenin. A Succinate increased protein levels of non-p-β-catenin and β-catenin in the mice kidney. *** P < 0.001, versus control group, n = 5. RAW 264.7 was treated with 500 μM succinate for 12 h. B Succinate enhanced protein levels of non-p-β-catenin and β-catenin in the RAW 264.7. *** P < 0.001, versus control group, n = 3, biologically repeated 3 times. C Succinate promoted translocation of non-p-β-catenin into the nucleus. ICG-001 (2 μM) pretreatment RAW 264.7 for 1 h, 500 μM succinate stimulation for 24 h and 48 h. D ICG-001 prevented the increase of CTGF mRNA induced by succinate. *** P < 0.001, versus control group, &&& P < 0.001, versus the succinate group, n = 3, biologically repeated 3 times. E The elevation of CTGF protein level was also lowered by ICG-001. *** P < 0.001, versus control group, &&& P < 0.001, versus the succinate group, n = 3, biologically repeated 3 times

Article Snippet: The cell supernatant was measured by mouse CTGF ELISA kit according to the manufacturer’s instructions (E-EL-M0340; Elabscience, Bethesda, MD).

Techniques: Expressing, Activation Assay, Control, Translocation Assay

Journal: iScience

Article Title: Microbiome assembly on Drosophila body surfaces benefits the flies to combat fungal infections

doi: 10.1016/j.isci.2022.104408

Figure Lengend Snippet: Bacterial load increase on the Drosophila body surfaces after eclosion for different times and variations in fly survival and AMP gene expressions (A and B) Time-dependent increase of the bacterial CFUs on the surfaces of female (A) and male (B) flies. Within each panel, different capital and lower letters labeled above each sample represent the significance of difference at the level of p < 0.01 and p < 0.05, respectively, after one-way ANOVA analysis. (C and D) Variation or non-variation of the CFU numbers formed on the LB medium with the addition of different antibiotics. Both the 2-DPE (C) and 10-DPE (D) flies were washed for CFU counting. DPE, days post eclosion. The antibiotic nafcillin largely inhibits the G+ bacteria whereas aztreonam inhibits the G- bacteria. The difference level of the two-tailed Student’s t test for significance is at: ∗∗∗, p < 0.001. (E) No obvious difference in the CFU numbers formed on different media. The 2-DPE flies were washed for CFU counting. Panels A-E: Values are represented as mean ± SD. (F and G) Survival of the different-age female flies against the topical infection of B. bassiana (F) and M. robertsii (G). Mocks one and two represent the two- and 10-DPE flies treated with 0.05% Tween 20, respectively. Plotted values are represented as mean ± SEM (SE of mean). More than 70 flies were used for each treatment and the experiments were repeated twice. (H and I) qRT-PCR analysis of the antimicrobial gene expressions after the topical infection of the two- and 10-DPE female flies with B. bassiana (Bb, panel H) and M. robertsii (Mr, panel I) for 48 h. The flies treated with 0.05% Tween 20 were used as mock controls. Values are mean ± SD. There were three replicates each with 10 flies, and the two-tailed Student’s t test was conducted to compare the expression level difference of AMP genes: ∗, p < 0.05; ∗∗, p < 0.01; ∗∗∗, p < 0.001; ∗∗∗∗, p < 0.0001. (See also Figure S1 and Table S1 ).

Article Snippet: To this end, each pooled bacterial sample was treated with 100 μl of Lysis Buffer (Mouse Tissue Direct PCR Kit, Tiangen Biotech, Beijing) for 30 min at 65°C and 5 min at 95°C.

Techniques: Labeling, Bacteria, Two Tailed Test, Infection, Quantitative RT-PCR, Expressing

Journal: iScience

Article Title: Microbiome assembly on Drosophila body surfaces benefits the flies to combat fungal infections

doi: 10.1016/j.isci.2022.104408

Figure Lengend Snippet:

Article Snippet: To this end, each pooled bacterial sample was treated with 100 μl of Lysis Buffer (Mouse Tissue Direct PCR Kit, Tiangen Biotech, Beijing) for 30 min at 65°C and 5 min at 95°C.

Techniques: Virus, Isolation, cDNA Synthesis, Lysis, Software, Electron Microscopy