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. 2016 Mar 22;5:e14052. doi: 10.7554/eLife.14052

Figure 4. Generation of a Prm1-mlCNBD-FRET transgenic mouse line.

Figure 4.

(A) Scheme of the Prm1-mlCNBD-FRET targeting vector. Expression of hemagglutinin(HA)-tagged mlCNBD-FRET is driven by the Protamine 1 promoter (Prm1); arrows indicate the position of genotyping primers (#1, 2). (B) Genotyping by PCR. In Prm1-mlCNBD-FRET mice, a 653 bp fragment is amplified (1). The targeting vector served as a positive control (+). (C) Western blot analysis of mlCNBD-FRET expression in lysates from different tissues from a transgenic male and a female. Lysates from HEK293 cells expressing mlCNBD-FRET served as positive control. Proteins have been labeled using an anti-GFP antibody. (D) Western blot analysis of mlCNBD-FRET expression in testis and sperm lysates from a wild-type and a transgenic male. Lysates from HEK cells expressing mlCNBD-FRET served as positive control. Proteins have been labeled using an anti-GFP antibody; tubulin has been used as a loading control. (E) Immunohistochemical analysis of mlCNBD-FRET expression in testis and sperm. Cryosections of mouse testis were probed with anti-HA antibody and fluorescent secondary antibody (purple); the fluorescence for cerulean (blue) or citrine (green) is also shown.

DOI: http://dx.doi.org/10.7554/eLife.14052.007

Figure 4—source data 1. Fertility parameters of mlCNBD-FRET transgenic males.
For matings, heterozygous males have been crossed with wild-type females. All data are represented as mean ± S.D., n numbers are indicated.
DOI: http://dx.doi.org/10.7554/eLife.14052.008
elife-14052-fig4-data1.docx (21.5KB, docx)
DOI: 10.7554/eLife.14052.008