c1q recombinant protein  (MedChemExpress)

Journal: Journal of Neuroinflammation

Article Title: Targeting microglial C1q alleviates blood–brain barrier disruption in the thalamus after cortical infarction

doi: 10.1186/s12974-026-03822-2

Figure Lengend Snippet: Single-cell RNA sequencing identifies microglia as the primary source of C1q in ipsilateral VPN of thalamus after dMCAO. A Scheme of scRNA-seq. B Umap of 11 major cell populations identified in the ipsilateral VPN in the Sham and dMCAO groups. Left: Integration of data from the Sham and dMCAO groups. Top middle: The Sham group. Bottom middle: The dMCAO group. Right: Stacked bar graphs of cell type proportions in the Sham and dMCAO groups ( n = 4 in each group). C Cell type annotation was performed on the basis of the expression of well-established marker genes. D Feature plots showing the expression of known cell markers. E Feature plots showing the expression of Tmem119, C1qa, C1qb and C1qc. F Co-stained with C1q (red) and Iba-1 (green), GFAP (green), NeuN (green) within ipsilateral VPN in Sham and dMCAO groups. Scale bar: 25 µm. G Umap of eight cell populations identified by microglial reclustering. H Proportions of all microglial subclusters in Sham and dMCAO groups. I Feature plots showing the expression of eight microglial subclusters

Article Snippet: Endothelial cells were treated with 1% DMSO, LPS, LPS-treated BV2 cell culture medium, C1q recombinant protein (Medchemexpress, Cat# HY-NP0194C), or C1q neutralizing antibody (GeneTex, Cat# GTX54404), separately.

Techniques: Single Cell, RNA Sequencing, Expressing, Marker, Staining

Journal: Journal of Neuroinflammation

Article Title: Targeting microglial C1q alleviates blood–brain barrier disruption in the thalamus after cortical infarction

doi: 10.1186/s12974-026-03822-2

Figure Lengend Snippet: Knockdown of microglial C1qa alleviates BBB disruption in the VPN of ipsilateral thalamus at 2 w after dMCAO. A The AAV vector maps and study design/timeline of left VPN of thalamus with AAV vectors carrying the CX3CR1 promoter. B Representative photomicrographs show the co-localization of GFP (green) and Iba-1 (red) in ipsilateral VPN at 2 w after dMCAO from animals with AAV- shC1qa injection. Scale bar: 25 μm. C Representative images show the co-localization of GFP (green) and C1q (red) in ipsilateral VPN at 2 w after dMCAO from animals with AAV injection. Scale bar: 25 μm. D Representative western blot showing the expression of C1qa in different AAV- shC1qa treatment sequence in Sham groups. Statistical analysis was performed using one-way ANOVA followed by Tamhane’s T2 test. Each bar represents the mean ± SD. * p < 0.05 vs AAV-shCon group at 2 w after dMCAO ( n = 4 in each group). E On day 14 post operation, the expressions of C1qa, ZO-1, OCLN, CLDN5, and VE-cad in ipsilateral VPN from Sham and dMCAO animals with AAV- shC1qa injection were showed through representative western blot. F - J On day 14 post operation, quantitative analyses of C1qa, ZO-1, OCLN, CLDN5, and VE-cad level relative to GAPDH in the ipsilateral VPN from Sham and dMCAO animals with AAV- shC1qa injection were conducted ( n = 4 in each group). Statistical analysis was performed using one-way ANOVA followed by Tamhane’s T2 test. (K) On day 14 post operation, representative images of extravascular albumin, IgG or fibrinogen deposits and CD8a positive cells in ipsilateral VPN from Sham and dMCAO animals with AAV- shC1qa injection were displayed. Scale bar: 25 μm. (L-O) On day 14 post operation, quantitative analyses of the area of albumin, IgG and fibrinogen leakage and the number of CD8a positive cells in the ipsilateral VPN from Sham and dMCAO animals with AAV- shC1qa injection were conducted ( n = 6 in each group). Statistical analysis was performed using Kruskal–Wallis H test. Each bar represents the mean ± SD. * p < 0.05 vs Sham + AAV-shCon group, # p < 0.05 vs dMCAO + AAV-shCon group, & p < 0.05 vs Sham + AAV- shC1qa group

Article Snippet: Endothelial cells were treated with 1% DMSO, LPS, LPS-treated BV2 cell culture medium, C1q recombinant protein (Medchemexpress, Cat# HY-NP0194C), or C1q neutralizing antibody (GeneTex, Cat# GTX54404), separately.

Techniques: Knockdown, Disruption, Plasmid Preparation, Injection, Western Blot, Expressing, Sequencing

Journal: Journal of Neuroinflammation

Article Title: Targeting microglial C1q alleviates blood–brain barrier disruption in the thalamus after cortical infarction

doi: 10.1186/s12974-026-03822-2

Figure Lengend Snippet: The interaction between microglia and endothelial cells and the activation of endothelial cells in the ipsilateral VPN of thalamus after dMCAO. A List of the molecular ligand-receptor pairing between microglia and endothelial cells from Sham and dMCAO groups. B Feature plots showing the expression of vWF and CD93. C Double-staining of Laminin (green) with CD93 (red) in the ipsilateral VPN in Sham and dMCAO groups. Scale bar: 25 μm. D Quantitative analyses of the expression of CD93 in Sham and dMCAO groups ( n = 6 in each group). Statistical analysis was performed using two-tailed t -test. E Representative western blot showing the expression of CD93 in VPN at 1–4 w after dMCAO. Statistical analysis was performed using one-way ANOVA followed by Tamhane’s T2 test. F Immunoprecipitation assay shows the levels of C1qa and CD93 in ipsilateral VPN in Sham and dMCAO groups ( n = 4 in each group). Statistical analysis was performed using two-tailed t -test. G Double-staining of C1q (red) with CD93 (green) in the ipsilateral VPN in Sham and dMCAO groups. Scale bar: 25 μm. H Representative western blot showing the expressions of VCAM-1 and ICAM-1 in VPN at 1–4 w after dMCAO. Quantitative analyses of VCAM-1 and ICAM-1 level relative to GAPDH ( n = 4 in each group). Statistical analysis was performed using one-way ANOVA followed by Tamhane’s T2 test. Each bar represents the mean ± SD. * p < 0.05 vs Sham group, # p < 0.05 vs contra. I Double-staining of VCAM-1 (green) with RECA-1 (red) in the ipsilateral VPN in Sham and dMCAO groups. Scale bar: 25 μm. J Colocalization analyses of VCAM-1 (green) and RECA-1 (red) in Sham and dMCAO groups

Article Snippet: Endothelial cells were treated with 1% DMSO, LPS, LPS-treated BV2 cell culture medium, C1q recombinant protein (Medchemexpress, Cat# HY-NP0194C), or C1q neutralizing antibody (GeneTex, Cat# GTX54404), separately.

Techniques: Activation Assay, Expressing, Double Staining, Two Tailed Test, Western Blot, Immunoprecipitation

Journal: Journal of Neuroinflammation

Article Title: Targeting microglial C1q alleviates blood–brain barrier disruption in the thalamus after cortical infarction

doi: 10.1186/s12974-026-03822-2

Figure Lengend Snippet: Microglial C1q activates endothelial cells via CD93 in the ipsilateral VPN of thalamus after dMCAO. A Representative images show the co-localization of RECA-1 (red) and VCAM-1 (grey) in ipsilateral VPN from Sham and dMCAO animals with AAV- shC1qa injection. Scale bar: 25 μm. B Immunoprecipitation assay shows the levels of C1qa and CD93 in ipsilateral VPN in Sham and dMCAO animals with AAV- shC1qa injection ( n = 4 in each group). Statistical analysis was performed using one-way ANOVA followed by Bonferroni test. C Representative western blot showing the expressions of VCAM-1 in ipsilateral VPN in Sham and dMCAO animals with AAV- shC1qa injection ( n = 4 in each group). Quantitative analysis of VCAM-1 level relative to GAPDH. Statistical analysis was performed using one-way ANOVA followed by Tamhane’s T2 test. * p < 0.05 vs Sham + AAV-shCon group, # p < 0.05 vs dMCAO + AAV-shCon group. D Representative western blot showing the expression of C1qa in BV2 cells with LPS treatment ( n = 4 in each group). Quantitative analysis of C1qa level relative to GAPDH. E Representative western blot showing the expression of VCAM-1 in brain endothelial cells with LPS treatment ( n = 4 in each group). Quantitative analysis of VCAM-1 level relative to GAPDH ( n = 4 in each group). Statistical analysis was performed using one-way ANOVA followed by Tamhane’s T2 test. Each bar represents the mean ± SD. * p < 0.05 vs Veh group. F Representative western blot showing the expression of VCAM-1 in each group ( n = 4 in each group). Quantitative analysis of VCAM-1 level relative to GAPDH. Each bar represents the mean ± SD. * p < 0.05 vs Veh group, # p < 0.05 vs BV2 or r-C1q groups. G Flow chart illustrates drug administration and experimental design. Rats received intracerebroventricular injection of DCBY02 or Veh at 3 days prior to dMCAO and every 2 days thereafter until the end of experiment. H – L Representative western blot showing the expressions of VCAM-1, ZO-1, OCLN, CLDN5, and VE-cad in ipsilateral VPN, respectively. Quantitative analyses of VCAM-1, ZO-1, OCLN, CLDN5, and VE-cad level relative to GAPDH from Sham and dMCAO animals with DCBY02 or Veh injection ( n = 4 in each group). Statistical analysis was performed using one-way ANOVA followed by Bonferroni test. Each bar represents the mean ± SD. * p < 0.05 vs Sham + Veh group, # p < 0.05 vs dMCAO + Veh group

Article Snippet: Endothelial cells were treated with 1% DMSO, LPS, LPS-treated BV2 cell culture medium, C1q recombinant protein (Medchemexpress, Cat# HY-NP0194C), or C1q neutralizing antibody (GeneTex, Cat# GTX54404), separately.

Techniques: Injection, Immunoprecipitation, Western Blot, Expressing

Journal: Journal of Neuroinflammation

Article Title: Targeting microglial C1q alleviates blood–brain barrier disruption in the thalamus after cortical infarction

doi: 10.1186/s12974-026-03822-2

Figure Lengend Snippet: Minocycline alleviates BBB disruption by downregulating microglial C1q-mediated VCAM-1 upregulation in ipsilateral VPN after dMCAO. A Binding conformation of minocycline to C1q. Left: overall view of minocycline in the binding position of C1q. Middle: three-dimensional (3D) close-up view of the binding conformation of minocycline on the C1q structure. Right: the enlarged image of the binding conformation of minocycline on the C1q structure. B Free energy distribution profile of the C1q-minocycline complex. C Experimental design/timeline of minocycline injection. D Representative western blot showing the expression of C1qa in different dose of minocycline treatment (+ : 10 mg/kg; + + : 20 mg/kg; + + + : 40 mg/kg, n = 3 in each group). Statistical analysis was performed using one-way ANOVA followed by Tamhane’s T2 test. Each bar represents the mean ± SD. * p < 0.05 vs Sham + Veh group, # p < 0.05 vs dMCAO + Veh group. E Representative western blot showing the expressions of C1qa, VCAM-1, ZO-1, OCLN, CLDN5, and VE-cad in ipsilateral VPN, respectively. F - K Quantitative analyses of C1qa, VCAM-1, ZO-1, OCLN, CLDN5, and VE-cad level relative to GAPDH from Sham and dMCAO animals with MC or Veh administration ( n = 4 in each group). Statistical analysis was performed using one-way ANOVA followed by Tamhane’s T2 test. L Representative images show the co-localization of Iba-1 (green), C1q (red) and DAPI (blue) in ipsilateral VPN from dMCAO animals with MC or Veh administration. Scale bar: 25 μm. M Quantitative analysis of Iba-1 + -C1q + cells from Sham and dMCAO animals with MC or Veh administration ( n = 6 in each group). Statistical analysis was performed using one-way ANOVA followed by Tamhane’s T2 test. N Representative images show the co-localization of VCAM-1 (green) and RECA-1 (red) in ipsilateral VPN from dMCAO animals with MC administration. Scale bar: 25 μm. O Representative microphotographs of immunostaining of NeuN, GFAP, and Iba-1 in ipsilateral VPN. Scale bar: 75 µm. (P-R) Quantitative analyses of NeuN positive cells ( P ), and GFAP ( Q ) and Iba-1 ( R ) positive densities were showed from ( O ) in ipsilateral VPN from Sham and dMCAO animals with MC or Veh administration ( n = 6 in each group). Statistical analysis was performed using one-way ANOVA followed by Bonferroni test. Each bar represents the mean ± SD. * p < 0.05 vs Sham + Veh group, # p < 0.05 vs dMCAO + Veh group, & p < 0.05 vs Sham + MC group

Article Snippet: Endothelial cells were treated with 1% DMSO, LPS, LPS-treated BV2 cell culture medium, C1q recombinant protein (Medchemexpress, Cat# HY-NP0194C), or C1q neutralizing antibody (GeneTex, Cat# GTX54404), separately.

Techniques: Disruption, Binding Assay, Injection, Western Blot, Expressing, Immunostaining

Journal: The Journal of Headache and Pain

Article Title: Inhibition of S100A4 decreases neurotoxic astrocyte reactivity and attenuates neuropathic pain via the TLR4/NF-κB pathway in a rat model of spinal nerve ligation

doi: 10.1186/s10194-025-02045-9

Figure Lengend Snippet: Exogenous S100A4 significantly contributes to A1 astrocyte activation in vitro. Western blot analysis revealed that GFAP and C3 protein levels were significantly increased after stimulation of 70% fusion primary astrocyte cultures with different concentrations of rat recombinant S100A4 protein ( A ), whereas 80 ng/ml S100A4 stimulation induced the greatest increase in GFAP and C3 expression ( B ). The data are presented as the means ± SEMs. * p < 0.05, ** p < 0.01, **** p < 0.0001, normal versus S100A4. ## p < 0.01, ### p < 0.001, #### p < 0.0001 versus 80 ng/ml S100A4. RT‒PCR analysis revealed changes in the mRNA levels of A1-specific astrocyte genes after 80 ng/ml S100A4 stimulation, similar to the effects of the commonly known “cytokine cocktail” containing TNF-α, IL-1α and C1q ( C ). The data are presented as the means ± SEMs. * p < 0.05, **** p < 0.0001 S100A4 versus normal. # p < 0.05, ### p < 0.001, #### p < 0.0001 TNF-α, IL-1α and C1q versus normal. Moreover, incubation with 80 ng/ml S100A4 resulted in higher OD values than those of normal cells and cells incubated with other concentrations of S100A4 ( D ). The data are presented as the means ± SEMs. **** p < 0.0001 versus 80 ng/ml S100A4. Immunofluorescence revealed that 70% of the fusion primary astrocytes (GFAP, green; DAPI, blue) incubated with TNF-α, IL-1α and c1q and S100A4 (80 ng/ml) for 24 h presented stronger GFAP (green) and C3 (red) fluorescence intensities and, accordingly, showed higher GFAP + C3 + fluorescence intensities ( E ) (scale bar, 100 μm). Quantitative analysis further confirmed these results ( F ). The data are presented as the means ± SEMs. **** p < 0.0001, normal versus S100A4. #### p < 0.0001, normal versus TNF-α, IL-1α and C1q

Article Snippet: Component C1q protein (MBS147399) was purchased from MedChemExpress (MCE) (Shanghai, China).

Techniques: Activation Assay, In Vitro, Western Blot, Recombinant, Expressing, Incubation, Immunofluorescence, Fluorescence