Fig. 4. BTZ obstructs NSC proliferation in the presence of growth factors and reduces Bcl-2/Bax ratio to induce apoptosis. (A) Images of neurospheres treated with 0.1% DMSO or 0.01, 0.1, 1, and 5 nM BTZ for 4 days in the presence of growth factors. Scale bar=250 μm. (B) The volume of neurospheres was estimated by measuring the diameter of individual neurospheres treated with DMSO or BTZ. (C, D) RT-PCR was performed to quantify mRNA level ratio of
Bcl-2 to
Bax. Total RNA was extracted from NSCs treated with DMSO or BTZ (0.1-5 nM) for (C) 24 h in proliferation media or (D) 48 h in differentiation media.
Gapdh was used as an internal control. (E, F) Representative images of protein bands of BCL-2 and BAX. Cell lysates were collected from NSCs treated with DMSO or BTZ (0.1-5 nM) for (E) 24 h in proliferation media or (F) 48 h in differentiation media, and a western blot analysis was conducted with BCL-2 and BAX antibodies. (G, H) Densitometer graphs of BCL-2/BAX protein expression. GAPDH was used as a loading control. Data are presented 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. 3.