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Fig. 6. GPR37 mediates HA signalling to stimulate mitosis. (A) HEK-T-REx-GPR37 cells were treated with and without doxycycline for 24 hours. Incubation with 2 nM HA for 1.7 hours led to an increase of cells in mitosis after induction of GPR37 expression. Immunostaining of cells with anti-phospho-histone H3 (1:1000) was used to determine cells in mitosis. 6x350 cells were counted, and the percentage of stained mitotic cells is given as means ± s.d. (B) Membrane currents were measured in the perforated patch configuration at a holding potential of -80 mV. Treatment with 1 nM HA induced an increase in membrane currents in COS-7 cells transiently expressing GPR37 (COS-GPR37), but not in mock-injected cells (COS-Mock). Membrane currents activated by HA in HEK-T-REx-GPR37 cells were blocked by application of 1 mM La³⁺ or 10 µM SKF. (C) Membrane-current densities were recorded from mock- and GPR37-transfected COS-7 cells. HA signal transduction was inhibited by pretreating cells for 2-3 hours with 200 ng ml^-1 pertussis toxin (PTX), for 30-60 minutes with 100 nM wortmannin (WORT), or 30 µM KN93 (KN). Each symbol represents one cell measured in the whole-cell (filled symbols) or the perforated patch (open symbols) configuration

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