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. 2020 Jun 2;11:2755. doi: 10.1038/s41467-020-16250-4

Fig. 6. FMRP(1–297)-tat is non-toxic to cultured Fmr1 KO granule cells.

Fig. 6

a–c Images of dual immunolabeled dissociated cultures of Fmr1 KO granule cells at 3 DIV for MAP2 and an anti-HA antibody 2 h following direct exposure to 500 nM (equivalent to 1.0 mg/kg in situ) HA-FMRP(1-297)-tat. Arrows denote position of representative cells labeling for both MAP2 and HA. Immunostaining was repeated with 3 different batches of cerebellar granule cell cultures. Scale bar, 25 µm. d–g Flow cytometric analysis of a live-dead cell viability assay in cerebellar granule cell cultures 24 h (d, e) and 5 days (f, g) after single exposure to the indicated concentrations of FMRP(1–297)-tat. The percentage of live cells in each cell population is not significantly different among the control group with no treatment, the vehicle treated group and cell groups treated with FMRP(1–297)-tat for 24 h (control, 99.4 ± 0.4%, n = 5; vehicle, 99.6 ± 0.2%, n = 5; 1 nM, 99.7 ± 0.1%, n = 3; 10 nM, 99.7 ± 0.1%, n = 4; 50 nM, 99.2 ± 0.4%, n = 4; 100 nM, 99.3 ± 0.3%, n = 5; 500 nM, 98.6 ± 1.1%, n = 3; F(6,28) = 0.763, p = 0.607, one-way ANOVA) and at 5 days (vehicle, 98.2 ± 1.3%, n = 4; 1 nM, 98.1 ± 1.6%, n = 4; 10 nM, 98.5 ± 1.2%, n = 4; 50 nM, 98.8 ± 0.9%, n = 4; 100 nM, 98.7 ± 0.6%, n = 4; 500 nM, 97.2 ± 1.0%, n = 3; F(6,27) = 0.406, p = 0.867, one-way ANOVA). N value represents the number of independent experiments repeated. Average values are mean ± s.e.m. Ctl Control, Veh vehicle. Source data are provided as a Source Data file.