Fig. 5

Knockdown of CTSS upregulated BDNF gene transcription and protein expression. A Clustering and heat map of the microarray analysis between the scramble siRNA and siCTSS groups of OEC-M1 cancer cells. The table indicates the mRNA fold change in siCTSS compared with scramble siRNA. Transient CTSS knockdown upregulated the mRNA expression of Bdnf (mRNA fold change level in the siCTSS group was normalized to that of the scramble siRNA group, fold-change cutoff ≥ 1.5 is shown in the table). B RJW-58 treatment enhanced Bdnf gene expression in HT-22 cells in a dose-dependent manner (N = 5). C Upregulation of Bdnf expression from 0 to 24 h after RJW-58 (2.5 µM) treatment (N = 6 per time window). The representative immunoblotting images of proBDNF and BDNF in (D) RJW-58 treated HT-22 cells after 48 h, and (E) the murine hippocampal tissues which were received 7-day consecutive RJW-58 dosing. F ELISA was used to validate the higher BDNF protein level expressing in the Ctss^−/− knockout mice hippocampal tissues than in that of Ctss^+/+ wildtype mice (N = 7). Bar charts indicate the mean ± SD. Asterisks indicate significant differences, Dunnett’s post-hoc test in B and C, unpaired two-tailed t-test in F, *p < 0.05 versus the DMSO group in B, versus the 0-h group in C, versus Ctss^+/+ mice in F