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. 2019 Jul 3;8:e45303. doi: 10.7554/eLife.45303

Figure 6. Requirement of Ror2 for adult neurogenesis.

(A–B) Co-immunoprecipitation Ror2 (A), not Ror1 (B), with Lrp4 in co-transfected HEK293T cell. (C–D) Increased Lrp4-Ror2 interaction in Agrin-treated HEK293T cell. (D) Quantitative analysis of data of C. Lrp4 intensity was normalized by that of IgG. Student’s t-test: t (4)=18.47, p<0.0001. (E) Increased Ror2 tyrosine phosphorylation in Agrin-treated neurosphere. Three independent experiments were performed. (F–G) Increased neurosphere size by Agrin and blockade by Lrp4 or Ror2 mutation. (F) Representative images. Scale bar, 100 µm. (G) Quantification of neurosphere size. One-way ANOVA test: F (5,314)=28.55, p<0.0001. Three independent experiments were performed. (H–I) Decreased Dcx+ cell density in Nes Ror2 CKO mice, compared with control. (H) Representative images. Scale bar, 100 µm. (I) Stereological quantification of Dcx+ cell density. n = 5 for each group. Student’s t-test: t (8)=4.523, p=0.0019. (J–L) Reduced BrdU+ cell density in Nes Ror2 CKO mice, compared with control. (J) Representative images. Scale bar 100 µm. (K) Stereological quantification of BrdU+ cell density. n = 5 for each group. Student’s t-test: t (8)=5.948, p=0.0003. (L) Similar density of SGZ Gfap+Sox2+ BrdU+ NSCs between the two genotypes. Student’s t-test: t (8)=1.22, p=0.2572. Data are mean ± s.e.m. **, p<0.01; ***, p<0.001.

Figure 6—source data 1. Requirement of Ror2 for adult neurogenesis.
elife-45303-fig6-data1.xlsx (13.6KB, xlsx)
DOI: 10.7554/eLife.45303.025
Figure 6—source data 2. Characterization of Ror2 mutant mice.
elife-45303-fig6-data2.xlsx (26.9KB, xlsx)
DOI: 10.7554/eLife.45303.026
Figure 6—source data 3. Characterization of inducible NSPCs-specific Ror2 knockout mice.
elife-45303-fig6-data3.xlsx (26.5KB, xlsx)
DOI: 10.7554/eLife.45303.027

Figure 6.

Figure 6—figure supplement 1. Generation and characterization of Ror2 mutant mice.

Figure 6—figure supplement 1.

(A) Generation of Ror2 conditional knockout mice. (B–C) Reduced Ror2 protein in Ror2 CKO hippocampus, compared with control. (B) Representative western blot. (C) Quantitative data of B. n = 3 for each group. Student’s t-test: t (4)=22.97, p<0.0001. (D) Similar brain gross morphology of Ror2 CKO and control mice. Scale bar, 5 mm. (E) Similar hippocampus morphology between control and Ror2 CKO mice. (F) Similar hippocampus volume of Ror2 CKO and control mice at P60. Student’s t-test: t (4)=0.416, p=0.6988. Scale bar, 200 µm. (G–H) Reduced density of Dcx+ cells in Ror2 CKO DG, compared with control. (G) Representative images. Scale bar, 50 µm. (H) Stereological. n = 4 for control and n = 3 for Ror2 CKO. Student’s t-test: t (5)=3.77, p=0.013. (I–J) Decreased numbers of BrdU+ cells in Ror2 CKO DG, compared with control. (I) Representative images. Scale bar 50 µm. (J) Stereological quantification. n = 4 for control and n = 3 for Ror2 CKO. Student’s t-test: t (5)=3.538, p=0.0166. Data are mean ± s.e.m. *, p<0.05; ***, p<0.001.
Figure 6—figure supplement 2. Generation and characterization of inducible NSPCs-specific Ror2 knockout mice.

Figure 6—figure supplement 2.

(A) Strategy of mouse crossing and Tam treatment. (B) Tam treatment diagram. (C) Reduced Ror2 protein in Nes Ror2 CKO DG after Tam treatment. n = 3 for each group. Student’s t-test: t (4)=7.7, p=0.0015. Data are mean ± s.e.m; **, p<0.01.