Fig. 2

Panx1-targeting nanobodies block Panx1 channel activity and show anti-inflammatory effects in vitro. a Panx1-targeting nanobodies (Nb1, Nb3 and Nb9) inhibit human Panx1 (hPanx1)-mediated extracellular release of ATP following potassium-induced channel opening in hPanx1-overexpressing DUBCA cells. Blocking effects of Nb1, Nb3, Nb9, non-targeting Nb (0–10,000 nM), carbenoxolone (CBX) (100 µM), lanthanum (La) (100 µM) and ¹⁰Panx1 (300 µM) were measured and expressed as the percentage of ATP relative to the release level triggered by potassium-enriched buffer (osmotic buffer) (n = 3 independent experiments). b Anti-inflammatory effects of Panx1-targeting nanobodies (Nb1, Nb3 and Nb9) were measured by analysing IL-1β signals in RAW264.7 cells. Anti-inflammatory effects of Nb1, Nb3, Nb9, non-targeting Nb (1000 nM) and dexamethasone (DEX) (100 µM) were measured and expressed as the percentage of IL-1β relative to the level triggered by LPS + ATP (n = 3 independent experiments). All data was analysed by parametric 1-way analysis of variance followed by post hoc tests with Bonferroni’s correction. Data were expressed as means ± S.D