Fig 1: Overexpression of Wnt2 in mPFC glutamatergic neurons alleviates myelin injury and improves cognition. Results of the a) Y‐maze and b) T‐maze tests showing the spontaneous alternation percentage in the control and Camk2a‐Wnt2 groups at 2 months after surgery. n = 15–16 per group. Results of the open field test showing c) the total distance travelled and d) time spent in the corner area and e) center area in the control and Camk2a‐Wnt2 groups at 2 months after surgery. n = 15–16 per group. f) Heatmaps generated from DTI axial views of FA acquired from the control and Camk2a‐Wnt2 groups at 2 months after surgery. g) Quantification of FA values in the corpus callosum in the control and Camk2a‐Wnt2 groups at 2 months after surgery. The measured values were normalized to the mean value of the control group. n = 11 per group. h) Representative images of Black‐gold staining and immunostaining of MBP and MAG in the corpus callosum in the control and Camk2a‐Wnt2 groups at 2 months after surgery. Scale bar: 200 µm. Quantification of i) Black‐gold staining and immunostaining of j) MBP and k) MAG in the corpus callosum in the control and Camk2a‐Wnt2 groups at 2 months after surgery. n = 4 per group. l) Representative TEM images of the corpus callosum in the control and Camk2a‐Wnt2 groups at 2 months after surgery. Scale bar: 1 µm. m) The percentage of myelinated axons in the corpus callosum in the control and Camk2a‐Wnt2 groups at 2 months after surgery. n = 4 per group. n) Scatterplots of the myelin g‐ratio as a function of the axon diameter in the corpus callosum in the control and Camk2a‐Wnt2 groups at 2 months after surgery. Axons were selected from 4 mice per group for measurement. n = 138 and n = 121 measured axons for each group, respectively. o) Representative immunostaining of CC1 and Olig2 in the corpus callosum in the control and Camk2a‐Wnt2 groups at 2 months after surgery. Scale bar: 50 µm. Density of p) CC1+Olig2+ cells and q) CC1−Olig2+ cells in the corpus callosum in the control and Camk2a‐Wnt2 groups at 2 months after surgery. n = 5 per group. r) Proportion of CC1+/Olig2+ cells in the corpus callosum in the control and Camk2a‐Wnt2 groups at 2 months after surgery. n = 5 per group. s) Immunoblot bands and t) quantification of MBP and u) MAG expression in OPCs in the control, T3, and T3+ rmWnt2 groups. The intensity of each immunoblot band was normalized to that of the β‐actin band. The measured values were normalized to the mean value of the control group. n = 3 per group. The data are presented as the mean ± SEM. p‐values were determined by Student's t‐test in (b–e), (i–k), (m), (p–r), and (t); by the Mann–Whitney test in (a), (g), and (n); and by 1‐way ANOVA with Tukey's post‐hoc analysis in (u). *p < 0.05, **p < 0.01, ***p < 0.001.
Fig 2: Secreted Wnt2 promotes cell proliferation in gastric cancer cell lines. A plasmid containing Wnt2 was stably transfected into CHO cells and CM containing secreted Wnt2 was collected for further study. (A) Expression of Wnt2 in CHO-W was confirmed by western blot analysis. (B) Quantification of the western blot analysis indicated that Wnt2 protein could be detected in CHO-W and the CM of CHO-W. Effects of normal growth media (DMEM + 10% fetal bovine serum), VecCM or VecWnt2CM on the growth of gastric cancer cells (C) SGC-7901 and (D) BGC823 were compared by an MTT assay. Data are presented as the mean ± standard deviation of three independent experiments. CM, conditioned medium; CHO-W, Wnt2-transfected CHO cells; CHO-V, empty vector-transfected CHO cells; DMEM, Dulbecco's modified Eagle's medium; VecCM, control CM; VecWnt2CM, CHO-Wnt2 CM; OD, optical density.
Fig 3: Expression of Wnt2 in human gastric cell lines. (A) Reverse transcription-quantitative polymerase chain reaction analysis of Wnt2 mRNA in four gastric cancer cell lines and a noncancerous cell line, GES-1. (B) Western blot analysis of Wnt2 protein in four gastric cancer cell lines and GES-1. The 40-kDa band was detected by anti-Wnt2 antibody (upper row), and the 42-kDa band was detected by anti-actin antibody (lower row) as the loading control.
Fig 4: Immunohistochemical staining of human gastric tissues. (A) Immunostaining of Wnt2 protein in human gastric tissues (magnification of the inset, ×20; main image, ×200). (Aa) Negative Wnt2 expression in normal gastric tissue. (Ab) Negative Wnt2 expression in non-tumor gastric mucosa. (Ac) Positive Wnt2 expression in gastric cancer tissue. (B) Expression level of Wnt2 protein in gastric cancer tissues was significantly higher compared with adjacent non-cancerous tissues. **P<0.001.
Fig 5: Effects of Wnt2 on wound healing in gastric cancer cells. Wound healing assay with (A) BGC823 and (B) gastric cancer cells. Microscopic observations were recorded at 0 and 48 h after scratching the cell surface. Representative images from each experiment are shown. Magnification, ×100. *P<0.05. NC, negative control.
Supplier Page from Abcam for Anti-Wnt2/IRP antibody [EPR3101(2)]