Biomolecules & Therapeutics : eISSN 2005-4483 / pISSN 1976-9148

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Fig. 2. BTZ increases TUBB3 expression in the absence of growth factors. (A-E) NSCs were treated with 0.1% DMSO or 0.01, 0.1, 1, and 5 nM BTZ in the absence of EGF and FGF2 for 4 days. (A, B) Immunofluorescence images of NSCs treated with (A) 0.1% DMSO or (B) 5 nM BTZ showing (a) nuclei stained with DAPI (blue), (b) neurons stained with TuJ1 (green), (c) astrocytes stained with GFAP (red), and (d) the merged images. Neurons are indicated by arrows (→). Scale bar=50 μm. (C-E) Quantification of (C) nuclei, (D) TuJ1- and (E) GFAP-positive cells. (F, G) RT-PCR was performed to quantify mRNA levels of Tubb3 and Gfap. Total RNA was extracted from NSCs treated with 0.1% DMSO or 0.1, 1, and 5 nM BTZ for 48 h in the absence of EGF and FGF2. Gapdh was used as an internal control. (H, I) Representative images of protein bands of (H) TUBB3 and (I) GFAP. After 48 h of treatment in the absence of EGF and FGF2, total cell lysates were collected, and a western blot analysis was conducted with TuJ1 and GFAP antibodies. GAPDH was used as a loading control. CNTF was used as a positive control for GFAP. Data are presented as the mean ± SEM (n=3). **p<0.01 (Student’s t-test). Uncropped images of western blots are shown in Supplementary Fig. 2.
Biomolecules & Therapeutics 2024;32:65~76 https://doi.org/10.4062/biomolther.2023.134
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