Fig. 8
Media cytokine and immune proteins match slice mRNA changes. OHSCs were treated with PLX3397 for 10 days in vitro (DIV) beginning on DIV4 to deplete microglia, followed by microglial repopulation in PLX3397-free medium for different durations. Media protein levels for glutamate, HMGB1, BDNF, and IL-4 were measured by ELISA. a Protein levels of the proinflammatory mediator HMGB1 was assessed by ELISA prior to depletion (DIV4), after depletion (DIV14) and after repopulation (DIV28). There was a significant treatment effect of microglial depletion and repopulation on media HMGB1 levels compared to untreated age-matched control cultures F(1,5) = 22.66, **p < 0.006. b Protein levels of the trophic Brain-derived neurotrophic factor (BDNF) were measured in culture media by ELISA after microglial repopulation. BDNF was increased by 28% in cultures that underwent microglial depletion and repopulation compared to age-matched untreated controls *p < 0.05, t test. c Media IL-4 protein was measured by ELISA after microglia repopulation. IL-4 showed a trend toward a significant increase after microglia repopulation (p = 0.07, t test). d OHSCs were grown in glutamate-free media, supplemented with l-glutamine (2 mM). Media glutamate was measured in OHSC media immediately after depletion (DIV14) and on repopulation day 14 (DIV 21) and 21 (DIV 28). Two separate experiments were analyzed at each time point. A transient increase in media glutamate was observed in the microglial depletion/repopulation group. ANOVA: F1,3 = 10.5, *p < 0.05