mab4101 sp c1q neutralizing antibody medchemexpress  (MedChemExpress)

Journal: Alzheimer's & Dementia

Article Title: Gut‐derived bacterial vesicles carrying lipopolysaccharide promote microglia‐mediated synaptic pruning

doi: 10.1002/alz.70331

Figure Lengend Snippet: bEVs induced C1q‐mediated synaptic pruning through activating Piezo1. (A and B) Western blot and quantitative analysis demonstrate the protein levels of C1q in HMC3 cells treated with PBS or bEVs ( N = 3 independent rounds of experimentation. In each round, three wells of HMC3 cells per group were measured and averaged to generate a single data point). (C) qPCR results and quantitative analysis demonstrate mRNA levels of C1q and C3 in HMC3 cells treated with PBS or bEVs ( N = 5 independent rounds of experimentation. In each round, four wells of HMC3 cells per group were measured and averaged to generate a single data point). (D and E) Western blot and quantitative analysis demonstrate protein levels of C1q in the cortex and hippocampus of 2‐month‐old mice treated with PBS or bEVs via tail vein injection for 24 h ( N = 6 and 8 mice for PBS and bEVs groups respectively). (F and G) qPCR results and quantitative analysis demonstrate mRNA levels of C1q and C3 in the cortex and hippocampus of 2‐month‐old mice treated with or without bEVs via tail vein injection for 24 h ( N = 3 and 5 mice for PBS and bEVs group, respectively). (H and I) Western blot and quantitative analysis demonstrate protein level of C1q in the hippocampus of 2‐month‐old mice treated with PBS, Pxb, bEVs, or bEVs+PxB via brain stereotactic injection for 24 h ( N = 3 mice in each group). (J and K) Representative immunofluorescence images and quantitative analysis show the presence of synaptic puncta (PSD95+) in a primary neuron and microglia cocultured system, and coculture cells were pretreated with ANX005 for 1 h, followed by treatment with PBS or bEVs ( N = 3 independent rounds of experimentation, each using primary neurons and microglia isolated from five different neonatal mice. In each round, six randomly captured neurons per well were measured and averaged to generate a single data point). (L and M) Western blot and quantitative analysis demonstrate protein levels of C1q in HMC3 cells pretreated with or without 5 µM GsMTx4 for 1 h, followed by bEV treatment ( N = 3 round of experimentation. In each round, four wells of HMC3 cells per group were measured and averaged to generate a single data point). Scale bar: 10 µm for J. Values are means ± SEM. ns, not significant; * p < .05. bEV, bacterial extracellular vesicle; PxB, polymyxin B.

Article Snippet: The cells were pretreated with 1 ng/mL ANX005 (HY‐P990545, MCE, USA), a humanized recombinant antibody against C1q, for 1 h to inhibit the complement cascade.

Techniques: Western Blot, Injection, Immunofluorescence, Isolation