Journal: Investigative ophthalmology & visual science

Article Title: The Effect of A2A Receptor Antagonist on Microglial Activation in Experimental Glaucoma.

doi: 10.1167/iovs.15-18024

Figure Lengend Snippet: FIGURE 5. Changes of microglial morphology and inflammatory mediators in the rat retinae with or without ZM241385 injection 2 weeks after the induction of COH. (A) The morphologic and quantity change of the microglia in different groups. In the COH rats, the amount of round or ameboid microglia in the retina decreased following intravitreal injection of ZM241385 (COHþZM) than that following the vehicle injection (COHþvehicle) and that without any injection (COH). The microglia extended 1 to 2 thick and short processes in the group of COHþZM, but still less ramified than that in the CTRL. White arrows pointed to microglia. Scale bar: 50 lm. (B) Changes of the mRNA expressions of TNF-a and IL-1b in retinae of the different groups. (C) Changes of the protein expressions of TNF-a, IL-1b, and Iba-1 in retinae of the different groups. *P < 0.05 represents comparison to CTRL group; #P < 0.05 represents comparison to COHþvehicle group. Each panel represents the average value. Error bar: mean 6 SD.

Article Snippet: Conditioned media collected from microglia treated with glutamate and ZM241385 for 24 hours was assayed for soluble TNF-a and IL-1b using ELISA kit (Wuhan Boster Biological Technology Ltd., Wuhan, Hubei, China) according to the manufacturer’s instructions.

Techniques: Injection, Comparison

Journal: Investigative ophthalmology & visual science

Article Title: The Effect of A2A Receptor Antagonist on Microglial Activation in Experimental Glaucoma.

doi: 10.1167/iovs.15-18024

Figure Lengend Snippet: FIGURE 7. Tumor necrosis factor-a and IL-1b secretion of the cultured microglia cells after treatment with glutamate and ZM241385. *P < 0.05 represents comparison to ctrl group; #P < 0.05 represents comparison to Glu1 group. Ctrl, control. Glu0.1, Glu1, Glu10 represent the culture media containing 0.1, 1, and 10 mM glutamate, respectively, Glu1þzm0.5 represents the culture media containing 1 mM glutamate and 0.5 lM ZM241385; zm0.5 represents the culture media containing 0.5lM ZM241385. Each panel represents the average. Error bar: mean 6 SD.

Article Snippet: Conditioned media collected from microglia treated with glutamate and ZM241385 for 24 hours was assayed for soluble TNF-a and IL-1b using ELISA kit (Wuhan Boster Biological Technology Ltd., Wuhan, Hubei, China) according to the manufacturer’s instructions.

Techniques: Cell Culture, Comparison, Control

Journal: Frontiers in Immunology

Article Title: Myeloid Vamp3 deletion attenuates CFA-induced inflammation and pain in mice via ameliorating macrophage infiltration and inflammatory cytokine production

doi: 10.3389/fimmu.2023.1239592

Figure Lengend Snippet: Myeloid Vamp3 deletion reduced inflammation and pain-related gene expression. (A-H) In comparison to WT mice, RT-qPCR showed the downregulation of IL-6 (A) , TNF-α (B) , CXCL11 (C) , TIMP-1 (D) , CD68 (E) , IL-1β (F) , COX-2 (G) , and CD54 (H) gene transcription levels in the ipsilateral plantar skins of Vamp3 Δmyel mice at day 14 after CFA injection. (I-K) CFA-injected paws of WT and Vamp3 Δmyel mice were dissected and homogenized in RIPA lysis and extraction buffer. The extracts were subjected to SDS-PAGE and Western Blotting. IL-1β (I) , COX-2 (J) , and CD54 (K) proteins were detected using their specific antibodies. As indicated, GAPDH or β-actin was probed as an internal loading control. (L) The ratios calculated for IL-1β, COX-2, and CD54 and the requisite internal standard (i.e. β-actin or GAPDH) for the CFA-injected paws of Vamp3 Δmyel mice were expressed relative to those for WT controls. Data are presented as mean ± SEM, n≥3 as indicated in the graph. *p<0.05, **p < 0.01, ***p < 0.001; unpaired two-tailed Student’s t-test, compared to the WT controls.

Article Snippet: The process was carried out in a similar manner, except for the use of specific antibodies: anti-IL-1β mouse monoclonal antibody (Servicebio, catalogue number GB122059-100, 1:500), anti-COX-2 rabbit polyclonal antibody (Servicebio, catalogue number GB11077-2-100, 1:1000), and anti-CD54 rabbit polyclonal antibody (Servicebio, catalogue number GB11106-100, 1:2000), respectively.

Techniques: Gene Expression, Comparison, Quantitative RT-PCR, Injection, Lysis, Extraction, SDS Page, Western Blot, Control, Two Tailed Test