Fig 1: BKCa openers control spasticity. (A) RNAseq expression of BKCa channel components in EA.hy926 cells. Inset represents the western blot of KCNMB4 in EA.hy926 cells, which was tested in four samples, and western blot of KCNMB4 expression in the spinal cord in control (n = 3) and spastic EAE spinal cord tissues (two shown, n = 6 total) showing an additional prominent band, not present in EA.hy926 cells, above the anticipated fragment size more notable during EAE. This antibody detected multiple bands in western blot analyses. (B) RNAseq expression of BKCa channel components in the spinal cords of spastic EAE (n = 3 mice) and age-matched normal mice (n = 3 mice). The results represent the mean ± SD. Inset represents the western blot of KCNMA1 in control and spastic EAE tissue (Antibody APC-021 detected a single band at the anticipated size and was repeated with antibody APC-151 with comparable results), and insets show a significant change in ratio between Kcnma1 and neurofilament light (Nefl) gene levels. (C) Percentage change in resistance to flexion of spastic hindlimbs in ABH mice in response to treatment with 5 mg·kg-1 p.o. VSN16R in water (n = 10 animals, n = 19 limbs) that were pretreated with saline or 1 mg·kg-1 i.p. paxilline (n = 7 animals, n = 14 limbs). (D) Percentage change in resistance to flexion of hindlimbs in ABH mice following injection i.p. with 1 mg·kg-1 paxilline (n = 7 animals, n = 13 limbs), 40 mg·kg-1 i.p. BMS-204352 (n = 7 animals, n = 13 limbs), 20 mg·kg-1 i.p. NS-1619 (n = 7 animals, n = 14 limbs) or 10 mg·kg-1 i.p. NS-11021 (n = 7 animals, n = 13 limbs). *Significant difference compared with baseline.