Journal: bioRxiv

Article Title: NINJ1 is activated by calcium-driven plasma membrane lipid scrambling during lytic cell death

doi: 10.1101/2024.10.23.619800

Figure Lengend Snippet: A, Stimulation of peritoneal macrophage with BzATP induces lytic cell death as assayed by LDH release into the supernatant. Pretreatment with A238079 prevents BzATP-induced cytotoxicity. * p < 0.05 by repeated measures one-way ANOVA with Geisser-Greenhouse correction and Tukey post-test from n = 3 independent experiments. B-C , YO-PRO-1 fluorescence was measured in primary mouse peritoneal macrophages at baseline, followed by stimulation with 300 µM BzATP over the course of 20 min. The leftmost graph in each panel indicates the average fluorescence intensity of YO-PRO-1 within cells over time (minutes). The middle graph depicts representative single cell tracings of YO-PRO-1 dye uptake. The rightmost graph (violin plot) in each panel indicates average YO-PRO-1 fluorescence intensity within individual cells (circles, squares) at 20 minutes following BzATP treatment. Peritoneal macrophages were stimulated with or without P2X7R perturbation by genetic knockout ( P2rx7 KO) ( B ) or pre-treatment with A438079 ( C ). D-E , Intracellular calcium measurements in primary mouse peritoneal macrophages before and after the addition of 300 µM BzATP. Graph indicates the ratio of fluorescence intensity of Fura-2 at 340 and 380 nm over time following BzATP treatment. Timepoints are shown relative to the first recorded measurement following BzATP treatment (t = 0 s). Peritoneal macrophages were stimulated with or without P2X7R perturbation by genetic knockout ( P2rx7 KO) ( D ) or pre-treatment with A438079 ( E ).

Article Snippet: R&D-AF5105) and P2X7 antibody (1:1000, Novus-NBP1-20180).

Techniques: Fluorescence, Knock-Out

Journal: Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie

Article Title: Antinociceptive and neuroprotective effect of echinacoside on peripheral neuropathic pain in mice through inhibiting P2X7R/FKN/CX3CR1 pathway.

doi: 10.1016/j.biopha.2023.115675

Figure Lengend Snippet: Fig. 2. Protocols of this study in vivo and vitro. (A) The major content of animal level test was to explore the anti-nociceptive and neuroprotective effect of ECH on CCI mice and to explore the potential mechanism primarily. (B) The major procedure of cell level test was to investigate the effect of ECH on BV2 cell after P2X7R activation induced by BzATP and LPS.

Article Snippet: These included a rabbit anti-IBA-1 N. Liu et al. Biomedicine & Pharmacotherapy 168 (2023) 115675 antibody (1:100 dilution, Protein-Tech, China), a rat anti-P2X7R antibody (1: 1000 dilution, Novus Biologicals, USA).

Techniques: In Vivo, Activation Assay

Journal: Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie

Article Title: Antinociceptive and neuroprotective effect of echinacoside on peripheral neuropathic pain in mice through inhibiting P2X7R/FKN/CX3CR1 pathway.

doi: 10.1016/j.biopha.2023.115675

Figure Lengend Snippet: Fig. 6. Effect of ECH on the expression of P2X7R in microglia of spinal cord dorsal horn in CCI mice. (A) Typical Immunofluorescence staining of Sham+NS, CCI + NS and CCI+ECH (200 mg/kg) group in spinal cord dorsal horn of mice. (B) Statistical chart of the number of co-expressed positive cells for P2X7R and IBA-1 in spinal cord dorsal horn of each group mice. Data are represented as the mean ± SD from six independent experiments (n = 6 per group). ##p < 0.01 compared with Sham+NS group; **p < 0.01compared with CCI+NS group. p Values are determined by one-way ANOVA and Tukey’s posttests.

Article Snippet: These included a rabbit anti-IBA-1 N. Liu et al. Biomedicine & Pharmacotherapy 168 (2023) 115675 antibody (1:100 dilution, Protein-Tech, China), a rat anti-P2X7R antibody (1: 1000 dilution, Novus Biologicals, USA).

Techniques: Expressing, Immunofluorescence, Staining