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Fig. 5. BTZ increases BDNF expression and phosphorylation of CREB in proliferating NSCs. (A, B) Representative images of the protein bands showing ubiquitin. NSCs were treated with 0.1% DMSO or 1 and 5 nM BTZ for 12 h (A) supplemented with growth factors for proliferation or (B) in the absence of growth factors for differentiation. After 12 h, total cell lysates were collected, and a western blot analysis was conducted with ubiquitin antibodies. Images of protein bands representing (C, D) BDNF and (E, F) p-CREB are shown. (G, H) Densitometer graphs of p-CREB expression. NSCs were treated with (C, E, G) 0.1% DMSO or BTZ (0.01-5 nM) for 24 h in proliferation media and (D, F, H) 0.1% DMSO or BTZ (0.1-5 nM) for 48 h in differentiation media. After treatment, cell lysates were progressed to western blot analysis with BDNF and p-CREB antibodies. GAPDH and total-CREB were used as loading controls. (I, J) RT-PCR was performed to quantify mRNA levels of Bdnf. Total RNA was extracted from NSCs treated with 0.1% DMSO or BTZ (0.1-5 nM) for (I) 24 h in proliferation media and (J) 48 h in differentiation media. Gapdh was used as an internal control. Data are shown as the mean ± SEM (n=3). *p<0.05, **p<0.01 (Student’s t-test). Uncropped images of western blots are shown in Supplementary Fig. 4, 5, and 6.
Biomolecules & Therapeutics 2024;32:65~76 https://doi.org/10.4062/biomolther.2023.134
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