Fig 2: Verification of the function of WDHD1 by animal experiments. (A) The transplanted tumor experiment showing that the inhibition of tumor volume by cisplatin in WDHD1 knockout group was more obvious than that in the control group. (B) A Western blot indicating that WDHD1 expression was upregulated in mice tissues implanted with A549/DDP cells, and MAPRE2 expression was increased in tumor tissues implanted with WDHD1 knockout cells. (C,D) The expression of Ki67 in the A549/DDP WDHD1 knockout group was significantly lower than that in the control group. The apoptotic index detected by a TUNEL assay was significantly higher in A549/DDP WDHD1 knockout group than that in the control group. ***p < 0.001; ****p < 0.0001. DDP: IC25 concentration for A549/DDP cells.

Fig 3: PCW-1001 reduced tumor growth of breast cancer cells in vivo. Female BALB/c nude mice were subcutaneously injected with 5 × 106 BT549 cells and treated with DMSO (Ctrl) or PCW-1001 (30 mg/kg) twice a week. (A–D) The tumor volume (A) and body weight (D) were measured periodically as indicated, and the tumor weights (B) and images (C) were obtained at the end of the experiment. (E) Cell lysates of the tumor were analyzed by immunoblotting with an anti-Ki67 antibody. ß-actin was used as the loading control. The data represent the results and are presented as mean ± standard deviation of three independent experiments. N, not significant; *P < 0.05, **P < 0.01.

Fig 4: Activation of GPR17 inhibited glioma formation.A U87MG and U251 cells treated with vehicle or MDL29951 (300 µM) for 48 h, and CCK-8 assay was performed to examine viable cell numbers. Data represent the means ± SEM from three independent experiments. B–D BALB/c nude mice with subcutaneously xenotransplanted tumor were intraperitoneally injected with 10 mg/kg MDL29951 for 16 days, and then tumor samples were collected for the measurements of tumor sizes (B), volumes (C), and weights (D), (n = 7). E H&E staining of sections from subcutaneously xenotransplanted tumors. Scale bar 10 µm. F, G Immunofluorescent staining against Ki67 (F) and TUNEL (G) of the tumor sections, scale bar 50 µm. H IHC staining against p-PKA or RNF2 of the subcutaneously xenotransplanted tumor. Scale bar 50 µm. I A diagram depicting the working model of the inhibitory effect of GPR17 on glioma tumorigenesis. For all panels, *p < 0.05, ***p < 0.001, Student’s t-test.

Fig 5: Effect of MTX and Dex on the number of immature neurons and cell proliferation. (A) The expression of DCX significantly decreases in the MTX-treated rats compared to the normal control rats; however, DCX expression significantly increases in the Dex co-treated groups when compared to the MTX-treated group, (B) Area-based % of Ki-67 immunohistochemical expression: Ki-67 expression is significantly lower in the MTX-treated group compared to the normal control group. Dex administration increases the % of Ki-67 positive cells when compared to the MTX-treated group. Ki-67 immunohistochemical staining of (C) normal control, (D) Dex control, (E) MTX, and (F) Dex co-treated group. Data are expressed as mean ± SD, n = 4 for Ki-67 immunohistochemical staining where n = 6 for DCX. (a) Significant difference from control, and (b) significant difference from MTX. Significance level is set at p < 0.05. One-way ANOVA with Tukey’s post hoc test were used in the statistical analysis. (C,F) Ki-67-stained sections 400×).