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A–D
Neurosphere (A, C) and XTT (B, D) assays were performed using mouse E14.5 primary neural progenitors transduced with a retroviral vector expressing Ttyh1 or short hairpin RNA specific to Ttyh1 (shTtyh1) (n = 4 for A and C, n = 3 for B and D).
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E
Ttyh1‐expressing cells in E15.5 embryonic brains that were intraventricularly electroporated at E13.5 were immunostained using an anti‐GFP primary (reporter gene) and Alexa Fluor 488‐conjugated secondary antibodies. GFP immunofluorescence (green) was merged with DAPI‐counterstained images (blue). VZ, ventricular zone; SVZ, subventricular zone; IZ, intermediate zone; CP, cortical plate. Scale bar: 100 μm.
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F
Quantification of (E) (n = 4).
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G
Double‐immunolabeling of E15.5 brain sections electroporated in utero with Ttyh1‐expressing plasmid at E13.5 using anti‐GFP (green) and Sox2 (red) antibodies, and Alexa Fluor 488‐ and 555‐conjugated secondary antibodies. Scale bar: 50 μm.
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H
Quantification of (G) (n = 3).
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I
Immunolabeling of E14.5 brain sections electroporated in utero with shTtyh1 alone or in combination with a shTtyh1‐resistant Ttyh1 (Ttyh1res) expression vector at E13.5 using anti‐GFP and Alexa Fluor 488‐conjugated secondary antibodies. The DAPI nuclear counterstain is shown in blue. Scale bar: 100 μm.
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J
Quantification of (I) (n = 3).
Data information: Error bars represent SD. Student's
‐test was used to determine statistical significance. *
0.001.
