Fig. 7

Trovafloxacin inhibits ATP release and migration in C5a-stimulated BV-2 cells. a BV-2 cells were stimulated with 10 nM of C5a in the presence or absence of Panx1 channel blockers, trovafloxacin (TVX, 1 μM), Brilliant Blue FCF (BBFCF, 5 μM), or ¹⁰Pnx1 (200 μM) for 30 min. Aliquots were collected from the medium every 10 min for ATP measurement. ATP was measured using the luciferin–luciferase assay. Values are represented as mean ± SEM. Percentage values were estimated with respect to basal ATP release before simulation at each condition. Each experiment was repeated at least three times in triplicates. Statistical difference was determined using linear regression with log(ATP) as the response. (p values calculated for treatment and treatment*time). *p < 0.05, C5a vs. control; **p < 0.05, C5a vs. TVX; ***p < 0.05, C5a vs. BBFCF, ****p < 0.05, C5a vs. ¹⁰Panx1. b Left panel shows representative images of BV-2 cells that were seeded in the upper compartment of the transwells for transmigration assay and treated with C5a in the absence or presence of TVX (1 μM) or BBFCF (5 μM). After 4 h, cells that had migrated to the bottom of the transwell were fixed, stained with crystal violet, and counted. The images are representative of the control treated with vehicle solutions, C5a, C5a + TVX, and C5a + BBFCF-treated cells. Graph corresponds to the quantification of transmigrated BV-2 cells after 4 h stimulation from images taken in five different fields at 20× per condition. Each experiment was repeated at least three times. One-way ANOVA with Tukey’s HSD test was used to measure statistical differences among groups. Values are expressed as mean ± SEM, *p < 0.05, control vs. C5a; **p < 0.05 C5a vs. C5a + TVX, or ***p < 0.05, C5a vs. C5a + BBFCF