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. 2020 Jan 15;13:561. doi: 10.3389/fncel.2019.00561

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Copyright © 2020 Cicvaric, Sachernegg, Stojanovic, Symmank, Smani, Moeslinger, Uhrin and Monje.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Lack of Podoplanin selectively decreases the survival of neuronal cells that are rescued by NGF treatment. Percentage of (A) neuronal and (B) glial cells derived from Pdpn^+/+ and Pdpn^−/− neurospheres surviving during 8 days (upper panels), showed a significant main effect of the genotype on number of surviving neurons (*P = 0.02, F_(1,10) = 8.69, n = 6) but not on the number of glial cells (P = 0.8, F_(1,8) = 0.07, n = 5). No significant effects of time were observed on a number of neuronal or glial cells. Percentage of (A) neuronal and (B) glial cells derived from Pdpn^+/+ and Pdpn^−/− neurospheres surviving after 4 days of NGF treatment, lower panels. In neuronal cells two-way ANOVA showed no significant effect of treatment (P = 0.2, F_(1,16) = 2.556, n = 6) but a significant effect of genotype (*P = 0.03, F_(1,16) = 5.84, n = 6) and a significant interaction between treatment and genotype (*P = 0.02, F_(1,16) = 7.07). Post hoc analysis showed a significant difference between Pdpn^+/+ and Pdpn^−/− without treatment (*P = 0.02) and between Pdpn^−/− with and without NGF treatment (*P = 0.059), as represented with asterisk on the graph. Analysis of glial cells showed a strong main significant effect of treatment (****P < 0.0001, F_(1,18) = 38.73, n = 5), significant main effect of the genotype (*P = 0.02, F_(1,18) = 7.695, n = 5) and a significant interaction between two factors (*P = 0.03, F_(1,18) = 5.942, n = 5). Post hoc analysis showed a significant difference between Pdpn^−/− with and without NGF treatment (****P < 0.0001) and between Pdpn^+/+ and Pdpn^−/− when treated with NGF (*P = 0.02), as represented with an asterisk on the graph. (C) Representative images of differentiated neurospheres immunolabeled with neuronal (MAP2) and glial (GFAP) markers (left and central pictures). In the right, upper and lower pictures, comprise representative images of immuno-based identified neurons (upper) and glial (lower) cells. Data are displayed as mean ± SEM.