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. Author manuscript; available in PMC: 2014 May 1.
Published in final edited form as: Mol Cell Neurosci. 2013 Jan 22;54:58–70. doi: 10.1016/j.mcn.2013.01.002

Figure 5. Exogenous expression of Pax6 impairs expressions of Tbr1 and Tbr2 in embryonic olfactory bulb.

Figure 5

(A, B) Sagittal sections of E14 OBs electroporated with pCAGEN (A) or pPax6 (B) together with pGFP. The sections were immunostained with anti-Tbr1 (Alexa 647, blue) and anti-Pax6 (Alexa 555, red) antibodies. GFP+ cells (Cy2, green) electroporated with pCAGEN are mostly localized in IZ and express Tbr1, but not Pax6 (A; closed-arrowheads). In contrast, many GFP+ cells electroporated with pPax6 are found in VZ. While all of them are Pax6+, some are Tbr1+ (B; closed-arrowheads). GFP+ cells that do not express Tbr1 are also found (B; open-arrowheads). Damaged region with a hole in the section shown in (A) is encircled with dotted lines. (C) Scatter plots showing percentages of Tbr1 expressing cells among GFP+ cells in E14 OB electroporated with pCAGEN (n=9) or pPax6 (n=9). The percentage is significantly decreased by pPax6 electroporation (***p<0.0005; unpaired t test). (D) Histogram of optical intensity (arbitrary unit) of Tbr1 in GFP+ cells electroporated with pCAGEN (blue) or pPax6 (red). Only GFP+ cells that were counted as Tbr1+ in (C) were used for the analysis. Trendlines were overlaid in the graph. (E, F) Similar to Tbr1 expression, Tbr2 (Alexa 555, red) is expressed by most of GFP+ cells (Cy2, green) electroporated with pCAGEN (E; closed-arrowheads), while both Tbr2+ (F; closed-arrowheads) and Tbr2– (F; open-arrowheads) cells are found among GFP+ cells electroporated with pPax6). (G) Scatter plots showing percentages of Tbr2 expressing cells among GFP+ cells in E14 OB elctroporated with pCAGEN (n=9) or pPax6 (n=9). The percentage is significantly decreased by pPax6 electroporation (***p<0.0005; unpaired t test). (H) Histogram of optical intensity (arbitrary unit) of Tbr2 in GFP+ cells electroporated with pCAGEN (blue) or pPax6 (red). Only GFP+ cells that were counted as Tbr2+ in (G) were used for the analysis. Trendlines were overlaid in the graph. Scale bars, 50 μm.