Fig. 5: IL-12p70 or IL-12p80 stimulation of murine embryonic primary myelinating culture results in reduced neurofilament and myelination.
a, Cell culture from E13 murine spinal cords treated with either IL-12p70 or IL-12p80 from DIV12 to DIV30. b, Immunocytochemistry from DIV30: blue, DAPI; white, MBP; pink, neurofilament. Scale bar, 100 µm. c, Quantification of neurofilament-covered area (%), biological replicates of vehicle-treated and IL-12p80-treated (n = 6) and IL-12p70-treated (n = 3) cell cultures, one-way ANOVA for vehicle control (mean = 6.780 ± s.e.m.), IL-12p70 (mean = 1.513 ± s.e.m.) and IL-12p80 (mean = 1.368 ± s.e.m.), df = 2, F = 13.09, P = 0.0010. d, Quantification of MBP-covered area (%), biological replicates of vehicle-treated and IL-12p80-treated (n = 6) and IL-12p70-treated (n = 3) cell cultures, one-way ANOVA for vehicle control (mean = 6.682 ± s.e.m.), IL-12p70 (mean = 1.237 ± s.e.m.) and IL-12p80 (mean = 1.295 ± s.e.m.), df = 2, F = 52.70, P < 0.0001. e, DAPI+ cell count, biological replicates of vehicle-treated and IL-12p80-treated (n = 6) and IL-12p70-treated (n = 3) cell cultures, one-way ANOVA for vehicle control (mean = 227.0 ± s.e.m.), IL-12p70 (mean = 104.0 ± s.e.m.) and IL-12p80 (mean = 110.2 ± s.e.m.), df = 2, F = 19.80, P = 0.0002. f, Quantification of apoptotic cells (%), biological replicates of vehicle-treated and IL-12p80-treated (n = 6) and IL-12p70-treated (n = 3) cell cultures, Kruskal–Wallis test for vehicle control (mean = 3.832 ± s.e.m.), IL-12p70 (mean = 7.333 ± s.e.m.) and IL-12p80 (mean = 12.50 ± s.e.m.), Kruskal–Wallis statistic = 11.35, P < 0.0001. g, Actin and pSTAT4 western blot analysis of IL-12p70-treated, IL-12p80-treated and non-treated primary oligodendrocytes; non-treated: 1.0 (left), IL-12p70-stimulated: 1.87 (center) and IL-12p80: 1.33 (right); pSTAT4 signal normalized to actin signal. df, degrees of freedom.
