Journal: Mediators of Inflammation

Article Title: Flemingia philippinensis Flavonoids Relieve Bone Erosion and Inflammatory Mediators in CIA Mice by Downregulating NF- κ B and MAPK Pathways

doi: 10.1155/2019/5790291

Figure Lengend Snippet: FPF treatment inhibited proinflammatory protein expression and AP-1 and STAT3 activation in the CIA mouse model. C57BL/6 mice were injected intradermally with CII at the base of the tail on day 0 and then challenged with 50 μ g CII subplantar at both hind limbs on day 18. On day 46, the paws and the spleen were dissected and total proteins were extracted ( n = 3-4 each group). Expressions of phosphorylated (p-) Jun D+c-Jun (a) and p-STAT3 (b) in paws and MCP-1 in the spleen (c) were determined using western blot and quantified by densitometric analyses. All data are mean ± standard error of the mean (SEM). ∗ P < 0.05 and ∗∗ P < 0.01 vs. the vehicle-treated control as determined using Student's t -test.

Article Snippet: Antibodies to the following proteins were purchased from Santa Cruz Biotechnology, Santa Cruz, CA: goat anti-mouse IgG2a-HRP (sc-2061), MMP-9 (sc-393859), cathepsin K (sc-48353), MCP-1 (sc-52701), phosphorylated STAT3 (sc-8059), phosphorylated I- κ B α (sc-8404), and phosphorylated NF- κ Bp65 (sc-101753); antibodies anti-Mouse IgG1-HRP, phosphorylated JNK1+JNK2+JNK3 (ab124956), phosphorylated p38 MAPK (ab178867), and phosphorylated Jun D+c-Jun (ab208035) were purchased from Abcam; phosphorylated ERK1/2 antibody (4370) was from Cell Signaling Technology, GAPDH antibody was from PeproTech Biotechnology, and ELISA kits for TNF- α , IL-10, IL-12/IL-23 p40 (total), and MCP-1 were from eBioscience.

Techniques: Expressing, Activation Assay, Injection, Western Blot

Journal: Scientific Reports

Article Title: Gut microbiota modulation induced by Zika virus infection in immunocompetent mice

doi: 10.1038/s41598-020-80893-y

Figure Lengend Snippet: Microbiota changes induced by ZIKV infection cause intestinal epithelium damage and intense leukocyte recruitment to the gut colon in adult immunocompetent mice. C57BL/6J mice (N = 5) were infected with ZIKV and colon tissue was collected after 14 days post-infection. The viral titer was assessed by qPCR in the serum ( A ) and the colon ( B ). The secretion of pro-inflammatory cytokines IL-12, TNF-α, IFN-γ and IL-1β, anti-inflammatory cytokines IL-10, and the cytokine IL-33, was measured by ELISA ( C ). The histological morphology of the colon was analyzed by HE staining and acquired in Zeiss microscope ( D ). Da-b represent section from uninfected mice and Dc-f represent sections from ZIKV infected mice. Db, Dd, and Df present zoomed area from Da, Dc, and De, respectively. Leukocyte infiltration and damaged epithelium are indicated by black arrows. Histological score comparing uninfected and ZIKV mice. The score represents the inflammatory cell infiltration ( E ) and epithelial changes ( F ). Statistical analyses were performed using a Student’s two-sample t-test, the non-parametric p-values were calculated with the Mann–Whitney test. Graphic bars represent a confidence interval of 95% (*p < 0.05; **p < 0.01 ***p < 0.001).

Article Snippet: The expression of IL-12, TNF-α, IFN-γ, IL-1β, IL-10, and IL-33 in the colon were assayed by ELISA according to the manufacturer’s instructions (Mouse TNF-α ELISA Ready-Set-Go, eBioscience, 88–7324–88; Mouse IL-1β ELISA Ready-SET-Go, eBioscience, 88–7013–88; Mouse IL-12/IL-23 total p40 ELISA Ready-SET-Go, eBioscience, 88–7120–88; Mouse IL-10 ELISA Ready-SET-Go 2° Generation, eBioscience, 88–7105–88; Mouse IL-33 DuoSet ELISA kit, R&D Systems, DY3626; Mouse IFN-gamma DuoSet ELISA kit, R&D Systems, DY485).

Techniques: Infection, Enzyme-linked Immunosorbent Assay, Staining, Microscopy, MANN-WHITNEY