Figure 1.
Co-staining of TLX cell type-specific markers in the SVZ.
A. Co-staining of TLX with Nestin in the SVZ of mouse brains. B. Co-localization of TLX with Mash1 in the SVZ of mouse brains. C. Co-staining of TLX with DCX in both the SVZ and the rostral migratory stream (RMS). LV stands for the lateral ventricles. Scale bar, 20 µm for all panels. Examples of TLX and Mash1 double-positive cells were indicated by arrows.
Figure 2.
Co-staining of TLX and BrdU in the SVZ.
Co-staining of TLX and BrdU in the SVZ of mice that were treated with BrdU for 1 week, followed by 4 week survival (long-term BrdU labeling), mice that were treated with BrdU once follwed by 6 hr survival (6 hr BrdU chase), or mice that were treated with BrdU once daily for 1 week (1 week BrdU treatment). LV stands for the lateral ventricles. Scale bar, 20 µm for all panels. Examples of TLX-positive, BrdU label-retaining cells that represent the ventricle-containing B1 cells were indicated by arrows, and examples of TLX-positive, BrdU label-retaining non-ventricle-containing B2 cells were indicated by arrowheads.
Figure 3.
Co-labeling of TLX with IdU and CIdU in the SVZ.
A. Schematics of IdU and CIdU treatment of adult mouse brains. B. Co-staining of TLX with IdU and CIdU in the SVZ of IdU and CIdU-treated mice. C. Co-staining of Ki67 with IdU and CIdU in the SVZ of IdU and CIdU-treated mice. Scale bar, 10 µm for all panels. The IdU, CIdU and Ki67 triple-positive cells were indicated by arrowheads.
Figure 4.
Reduced neural progenitor populations in the SVZ of TLX−/− brains.
A. short-term (6 hr pulse) BrdU labeling along with Mash1 and TLX staining in the SVZ of wild type (WT) and TLX−/− brains. The top panels show Mash1 single staining and the bottom panels show merged images of Mash1, brdU and TLX triple staining. B. Quantification of Mash1+ cells in the SVZ of WT and TLX−/− brains. Data are represented as means ± s.d. *p<0.001 by Student's t-test. C. Quantification of Mash1+Ki67+ cells from Mash1+ cells in the SVZ of WT and TLX−/− brains. Data are represented as means ± s.d. *p<0.001 by Student's t-test. D. DCX and TLX staining in the SVZ of WT and TLX−/− brains. The top panels show DCX single staining and the bottom panels show merged images of DCX and TLX double staining. Scale bar, 20 µm for all panels.
Figure 5.
Reduced BrdU label-retaining cells and increased GFAP-positive cells in the SVZ of TLX−/− brains.
A. There are reduced numbers of total cells and BrdU label-retaining cells in the SVZ of TLX−/− brains as revealed by Dapi staining (blue) and BrdU label (green) -retaining, and increased GFAP-positive cells as revealed by GFPA staining (purple). Both wild type (WT) and the TLX−/− mice were treated with BrdU once daily for 1 week, followed by 4 week survival. B. Quantification of Dapi-positive cells in the SVZ of wild type (WT) and TLX−/− brains. *p = 0.0015 by Student's t-test, n = 3. C. Quantification of BrdU label-retaining cells in the SVZ of WT and TLX−/− brains. *p = 0.019 by Student's t-test, n = 3. Error bars are standard deviation of the mean. Scale bar, 20 µm for all panels.
Figure 6.
Whole mount staining revealed increased GFAP staining and scar-like GFAP-positive signals in the SVZ of TLX−/− brains.
A. GFAP staining in the SVZ of TLX+/− and TLX−/− brains. Dapi and Vimentin (vim) staining was included as counter staining. B. Images of higher magnification of vimentin (1), GFAP(2), Dapi (3), and merged (4) staining in the SVZ of TLX−/− brains. Scale bar, 20 µm for all panels. Loss of Dapi staining in the Scar-like GFAP+ foci was indicated by asterisks.