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. 2020 Oct 14;10(10):1440. doi: 10.3390/biom10101440

Figure 1.

Figure 1

Erinacine C induces NGF expression in 1321N1 astrocytoma cells. (A) PC12 cells after 48 h of incubation in medium supplemented with (a) 0.5% EtOH, (b) 200 ng/mL of recombinant NGF, (c) 5 µg/mL erinacine C, (d) or maintained in medium conditioned by 1321N1 cells that were incubated with 5 µg/mL erinacine C. Neurite-like membrane protrusions as signs of PC12 differentiation are marked by white arrows. (B) PCR fragments after gel electrophoresis of semiquantitative RT-PCR samples to compare regulation of ngf expression in 1321N1 cells to the housekeeping gene gapdh under normal, 0.5% EtOH, and 5 µg/mL erinacine C conditions 48 h after onset of compound treatment. (C) Relative expression levels of ngf 1312N1 cells, EtOH controls and erinacine C exposed cells (n = 3). (D) PC12 cells transfected with pCS-dnTrkA-Citrine (green fluorescence) treated with 0.5% EtOH, 200 ng/mL recombinant NGF or conditioned medium of 1321N1 cells incubated in 5 µg/mL erinacine C. While nonfluorescent and pCS-FynmCitrine transfected control cells are marked by black arrows, green fluorescent cells with impaired TrkA-signaling are marked by white arrows. The differentiation of PC12 cells was quantified regarding (E) the percentage of cells displaying membrane protrusions (n = 3 independent cultures) and (F) the average length of neurite-like structures established by differentiating cells (n = 180). * p < 0.05, ** p < 0.01, **** p < 0.0001. Scale bar: 100 µm.