Fig. 8
TNFRSF12A in both AD and GBM. (a) Relative mRNA expression levels of Wnt pathway genes in GBM cell lines (G35 and 84) following successful TNFRSF12A knockdown. Non-transfected GBM cells lines were used as the mock group (Mock), while cells transfected with negative siRNA served as the negative control (NC). Results represent data from three separate experiments. Data are presented as mean ± standard deviation (SD) (n = 3 biological replicates). Significance levels were determined using two-way ANOVA with Bonferroni’s post-hoc test (*P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001). (b) Western blot analysis (top panel) and quantification (bottom panel) of TWEAK protein expression in SH-SY5Y cells with or without TNFRSF12A-siRNA. Band intensities were normalized to ponceau (total protein) for each sample and to the average of the control group (Mock). Non-transfected SH-SY5Y cells were used as the mock group (Mock), while cells transfected with negative siRNA served as the negative control (NC). Representative images are shown. Data are presented as mean ± SD (n = 3 biological replicates). Statistical significance was assessed using two-way ANOVA with Bonferroni’s post-hoc test (P < 0.05; ns, no significance). (c, d) Western blot analysis (left) and quantification (right) of full-length amyloid precursor protein (APP-FL) and its C-terminal fragment (APP-CTF) in SH-SY5Y cells with or without TNFRSF12A-siRNA. Band intensities were normalized to ponceau (total protein) and to the Mock group’s average. Non-transfected SH-SY5Y cells were used as Mock, while cells transfected with negative siRNA served as NC. Representative images are shown. Data are presented as mean ± SEM (n = 3). Statistical significance was determined using one-way ANOVA with Bonferroni’s post-hoc test (P < 0.05; ns, no significance). (e) Relative mRNA expression levels of APP gene in the SH-SY5Y cell line following successful TNFRSF12A knockdown. Data are presented as mean ± SD (n = 6). Statistical significance was determined using an unpaired Student’s t-test (P < 0.05). (f) Schematic diagram of the mechanism of TNFRSF12A in GBM and AD.
