Fig. 4

Glial activation and VZ disruption in the IVH in vitro model. a, b Comparisons of the expression of GFAP and βIV tubulin at day 7 after 48 h of control (a) or blood (b). a shows a qualitatively higher expression of βIV (arrows) and a lower expression of GFAP when compared to blood (b). a′, b′ High magnification (×63) of a, b in which a′ shows several multiciliated EC (asterisk) vs b′ that shows a lack of multiciliated EC and changes in the morphology of the VZ cells with the increase of GFAP projections (arrowhead), as well as dislocation (asterisk) and diminution of the expression of N-Cad. c Graphical representation of the roundness of GFAP-labeled cells after 48 h of treatment (control vs. blood), In which the cells are significantly less round in blood conditions (p < 0.001). Data are mean of the roundness of 2 different experiments with at least n = 10 d GFAP protein levels quantified by WB normalized with β actin after 48 h of treatment, in which blood exposure increases the expression of GFAP. Data are mean from a single experiment with n = 8. e Comparisons of the expression of EMA at day 5 after 3 h of treatment at day 6 after 24 h of treatment and at day 7 after 48 h of treatment. The immunocytochemistry shows a decrease in the expression of EMA at 24 and 48 h after blood induction. f Quantification of EMA-positive cells. After 24 and 48 h treatment the mean percentage of cells expressing EMA was significantly decreased. Data are mean of the percentage of cells expressing EMA, from 2 experiments with at least n = 5 wells at 3, 24, and 48 hours, respectively (***p < 0.001; Student t-test). Scale bars: a, b = 50 μm; a′, b′ = 10 μm; e = 20 μm