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. 2025 Jan 9;26(2):493. doi: 10.3390/ijms26020493

Figure 4.

Figure 4

Induction of differentiation in glioblastoma cell C6 and neuroblastoma cell IMR-32 by TEG (0.1%) and Wi-N (5 µM). (A) Phase contrast images showing astrocytic and neuron-like morphology in TEG- and Wi-N-treated C6 (30 days, left side) and IMR-32 (45 days, right side) cells, respectively. Images were captured at 10X magnification. (B,C) Western blotting analysis showing increased levels of glial differentiation proteins GFAP, MAP2, and PSD-95 in treated C6 cells (B) and increased levels of neurodifferentiation proteins NeuN, GAP-43, NF200 in treated IMR-32 cells (C). Quantification from three independent experiments is shown on the right side. (D) Immunostaining of GFAP (in C6) and NeuN (in IMR-32) showed increased fluorescence intensity in treated cells. The scale bars represent 10 µm in images of C6 cells and 20 µm in images of IMR-32 cells, respectively. Quantification from three independent experiments is shown below the images. (E) RT-qPCR analysis shows upregulated GFAP and MAP2 expression in treated C6 cells and upregulated GAP-43, NF200, and NeuN in treated IMR-32 cells. All data were normalized against the control group and plotted as fold difference (mean ± SD, n = 3). ns p ≥ 0.05, * p < 0.05, ** p < 0.01, *** p < 0.001 denote statistical significant differences from the control group (one-way ANOVA with Dunnett’s multiple comparisons). Control: 0.05% DMSO; ns: not significant; TEG: triethylene glycol; Wi-N: Withanone; Wi-A: Withaferin A.