rat anti c3ar  (Hycult Biotech)

Journal: Theranostics

Article Title: Microglia exacerbate white matter injury via complement C3/C3aR pathway after hypoperfusion

doi: 10.7150/thno.35841

Figure Lengend Snippet: Activation of complement C3-C3aR/ITGAM pathway in BCCAO rats. A Quantitative RT-PCR analysis of the expression of C1qa, C1qb, C4b, C3 , C3ar , and Itgam in the striatum of control and BCCAO rats at day 7, 14, and 28 after BCCAO surgery. The values are normalized to those of the control group. n = 3-7 in each group. B Western blots and quantification for C3, C3aR, ITGAM, and β-actin in the striatum of control and BCCAO rats at day 7, 14, and 28 after surgery. C Representative images and quantification of complement C3 puncta (red) deposition on myelin (MBP + , green) in the striatum of BCCAO and control rats. n = 3-4 animals in each group at day 7, 14, and 28 after surgery. Scale bar=10 μm. The data are shown as the mean ± SD. ***, p < 0.001; **, p < 0.01; *, p < 0.05; NS, not significant; the BCCAO group vs. the control group.

Article Snippet: Rat anti-MBP (Abcam, ab7349, 1:400), mouse anti-SMI32 (Biolegend, San Diego, CA, 801701, 1:100), rabbit anti-APC (Abcam, ab72040, 1:50), rabbit anti-Iba-1 (Wako, Richmond, VA, 019-19741, 1:300), rabbit anti-CD86 (Abcam, ab112490, 1:200), mouse anti-CD68 (AbD Serotec, MCA341, 1:200), rabbit anti-C3 (Abcam, ab200999, 1:200), mouse anti-ITGAM (AbD Serotec, Oxford, UK, MCA618, 1:100), rat anti-C3aR (Hycult Biotech, Uden, Netherlands, HM1123, 1:100) antibodies were used.

Techniques: Activation Assay, Quantitative RT-PCR, Expressing, Western Blot

Journal: Theranostics

Article Title: Microglia exacerbate white matter injury via complement C3/C3aR pathway after hypoperfusion

doi: 10.7150/thno.35841

Figure Lengend Snippet: Genetic deletion of C3ar1 attenuates microglia activation and reverses white matter injury in BCAS mice. A Representative images and quantification of C3aR (green) and Iba-1 (red) double-positive cells and microglia cells (Iba-1 + cells, red) in the striatum of WT, C3aR-KO, BCAS, and BCAS/C3aR-KO mice at day 28 after surgery. Scale bar=25 μm. B Western blots and quantification for CD86, iNOS, MBP and β-actin in the striatum of WT, C3aR-KO, BCAS, and BCAS/C3aR-KO mice at day 28 after surgery. C Representative images and quantification of damaged axon (SMI32 + , red) relative to myelin (MBP + , green) and mature oligodendrocyte (APC + cells, red) in the striatum of WT, C3aR-KO, BCAS, and BCAS/C3aR-KO mice at day 28 after surgery. Scale bar=50 μm. D Western blots of total- and phospho-STAT3 (pSTAT3) and β-actin in the striatum of WT, C3aR-KO, BCAS, and BCAS/C3aR-KO mice at day 28 after surgery. E and F Quantification of total STAT3/ β-actin ( E ) and phospho-STAT3/STAT3 ( F ) levels in the striatum of WT, C3aR-KO, BCAS, and BCAS/C3aR-KO mice at day 28 after surgery. The data are shown as the mean ± SD. n = 3-4 animals in each genotype group. ***, p < 0.001; **, p < 0.01; *, p < 0.05; NS, not significant.

Article Snippet: Rat anti-MBP (Abcam, ab7349, 1:400), mouse anti-SMI32 (Biolegend, San Diego, CA, 801701, 1:100), rabbit anti-APC (Abcam, ab72040, 1:50), rabbit anti-Iba-1 (Wako, Richmond, VA, 019-19741, 1:300), rabbit anti-CD86 (Abcam, ab112490, 1:200), mouse anti-CD68 (AbD Serotec, MCA341, 1:200), rabbit anti-C3 (Abcam, ab200999, 1:200), mouse anti-ITGAM (AbD Serotec, Oxford, UK, MCA618, 1:100), rat anti-C3aR (Hycult Biotech, Uden, Netherlands, HM1123, 1:100) antibodies were used.

Techniques: Activation Assay, Western Blot

Journal: Theranostics

Article Title: Microglia exacerbate white matter injury via complement C3/C3aR pathway after hypoperfusion

doi: 10.7150/thno.35841

Figure Lengend Snippet: C3aR inhibition suppresses microglial activation and microglia redistribution to myelin in BCCAO rats. A Western blots and quantification for CD86, iNOS, and β-actin in the striatum of the sham+vehicle, sham+C3aR antagonist, BCCAO+vehicle, and BCCAO+C3aR antagonist groups. n = 3-6 in each group. B Representative images of microglia cells (Iba-1 + cells, red) contacting with myelin (MBP + , green) in the striatum of sham+vehicle, sham+C3aR antagonist, BCCAO+vehicle, and BCCAO+C3aR antagonist groups. Scare bar=50 μm. n= 3-4 in each group. C-D Quantification of the proportion of microglia cells adhered to myelin relative to the number of myelin fibers ( C ) and the number of microglia cells ( D ). The data are shown as the mean ± SD. ***, p < 0.001; **, p < 0.01; *, p < 0.05; NS, not significant; the BCCAO group vs. control group. C3aRA: C3aR antagonist.

Article Snippet: Rat anti-MBP (Abcam, ab7349, 1:400), mouse anti-SMI32 (Biolegend, San Diego, CA, 801701, 1:100), rabbit anti-APC (Abcam, ab72040, 1:50), rabbit anti-Iba-1 (Wako, Richmond, VA, 019-19741, 1:300), rabbit anti-CD86 (Abcam, ab112490, 1:200), mouse anti-CD68 (AbD Serotec, MCA341, 1:200), rabbit anti-C3 (Abcam, ab200999, 1:200), mouse anti-ITGAM (AbD Serotec, Oxford, UK, MCA618, 1:100), rat anti-C3aR (Hycult Biotech, Uden, Netherlands, HM1123, 1:100) antibodies were used.

Techniques: Inhibition, Activation Assay, Western Blot

Journal: Theranostics

Article Title: Microglia exacerbate white matter injury via complement C3/C3aR pathway after hypoperfusion

doi: 10.7150/thno.35841

Figure Lengend Snippet: C3aR inhibition prevents behavioral deficits and white matter injury in BCCAO rats. A Five-day spatial learning performance measured as the latency to reach the platform in the Morris water maze test in sham+vehicle, sham+C3aR antagonist, BCCAO+vehicle, and BCCAO+C3aR antagonist groups. n= 12-15 animals for each group. B Spatial memory performance measured as the number of entries into the platform quadrant and the percentage of time spent in the platform quadrant in the Morris water maze test in the sham+vehicle, sham+C3aR antagonist, BCCAO+vehicle, and BCCAO+C3aR antagonist groups, n = 12-15 animals in each group. C Spatial memory performance measured as discrimination time in the new object recognition test in the sham+vehicle, sham+C3aR antagonist, BCCAO+vehicle, and BCCAO+C3aR antagonist groups. n = 4-6 animals for each group. D Western blots and quantification for myelin basic protein (MBP) and β-actin in the striatum of sham+vehicle, sham+C3aR antagonist, BCCAO+vehicle, and BCCAO+C3aR antagonist groups. E - F Representative images and quantification of damaged axon (SMI32 + , red) relative to myelin (MBP + , green) ( E ) and mature oligodendrocyte (APC + cells, red) ( F ) in the striatum of sham+vehicle, sham+C3aR antagonist, BCCAO+vehicle, and BCCAO+C3aR antagonist groups. n = 3-4 animals in each group. Scale bar=50 μm. The data are shown as the mean ± SD. **, p < 0.01; *, p < 0.05; the BCCAO group vs. control group. C3aRA: C3aR antagonist.

Article Snippet: Rat anti-MBP (Abcam, ab7349, 1:400), mouse anti-SMI32 (Biolegend, San Diego, CA, 801701, 1:100), rabbit anti-APC (Abcam, ab72040, 1:50), rabbit anti-Iba-1 (Wako, Richmond, VA, 019-19741, 1:300), rabbit anti-CD86 (Abcam, ab112490, 1:200), mouse anti-CD68 (AbD Serotec, MCA341, 1:200), rabbit anti-C3 (Abcam, ab200999, 1:200), mouse anti-ITGAM (AbD Serotec, Oxford, UK, MCA618, 1:100), rat anti-C3aR (Hycult Biotech, Uden, Netherlands, HM1123, 1:100) antibodies were used.

Techniques: Inhibition, Western Blot

Journal: Acta Neuropathologica Communications

Article Title: Protective effects of decay-accelerating factor on blast-induced neurotrauma in rats

doi: 10.1186/2051-5960-1-52

Figure Lengend Snippet: DAF treatment reduces interaction of C3a-C3aR in the rat brain tissue after blast exposure. Representative photomicrographs of C3a-C3aR interaction in frontal grey matter ( a ) and hippocampus (DG) ( b ) of frozen sections stained with anti-C3a (red) and anti-C3aR (green) antibodies. Original magnification of × 200 (grey matter) and × 400 (DG). Scale bars, 200 μm (grey matter) and 100 μm (DG). n = 8 for control, 3 and 24 h experimental groups. n = 5 for 48 h experimental groups.

Article Snippet: Mouse anti-rat C3a receptor (C3aR) and mouse anti-rat C5b-9 antibodies were acquired from Hycult Biotech Inc (Plymouth Meeting, PA).

Techniques: Staining