XB-IMG-157817
Xenbase Image ID: 157817
|
Additional file 3: Both SIX1 and SIX4 are expressed in DRG. (A) Specificities of anti-SIX1 and anti-SIX4 antibodies are validated by using Six1 or Six4 single homozygous knockout mice, Six1 -/- or Six4 -/- . To evaluate the specificities of the antibodies, the trigeminal ganglia (Vg) are subjected to immunofluorescence staining, in which both SIX1 and SIX4 are expressed during the development [48]. The rat polyclonal antibody against mouse SIX1 [48] detects SIX1 protein in Six4 -/- embryo (denoted as Six4 LacZ/LacZ ) [50], but not in Six1 -/- embryo (denoted as Six1 EGFP/EGFP ) [41]. The guinea pig polyclonal antibody against mouse SIX4 [39] recognizes SIX4 in Six1 EGFP/EGFP embryo, but none in Six4 LacZ/LacZ . These results show lack of cross-reactivity of anti-SIX1 and anti-SIX4 antibodies with SIX4 and SIX1, respectively. Bottom line shows merged images; SIX1 and SIX4 in wild type, EGFP expressed from Six1 knockout alleles and SIX4 in Six1 EGFP/EGFP , SIX1 and β-galactosidase expressed from Six4 knockout alleles in Six4 LacZ/LacZ . Scale bars: 50 μm. (B) Similar distribution of SIX1 and SIX4 in developing mouse DRG. In E11.5 mouse embryo, the majority of SIX1-positive-cells in DRG (green) are labeled with SIX4 immunofluorescence (magenta), as shown in the merged panel. The relative intensities of immunofluorescent signals for SIX1 and SIX4 vary among DRG neurons. Dashed lines demarcate the position of the ectoderm and spinal cord (sc). Scale bar: 50 μm. (TIFF 6 MB) Image published in: Yajima H et al. (2014) Copyright © 2014 Yajima et al. This image is reproduced with permission of the journal and the copyright holder. This is an open-access article distributed under the terms of the Creative Commons Attribution license Larger Image Printer Friendly View |