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. 2017 Nov 20;293(1):148–162. doi: 10.1074/jbc.M117.813931

Figure 5.

Figure 5.

Tamoxifen-induced deletion of Fhod3 in the adult heart does not lead to lethality. A, PCR analyses of genomic DNAs from hearts and kidneys of TAM-treated Fhod3 iKO (Fhod3flox/−;MerCreMer+) and control littermate (Fhod3flox/−;MerCreMer−) mice. The PCR product from the wild-type allele (Fhod3+) or the null allele (Fhod3−) was 329 bp in length. The PCR product from the floxed allele (Fhod3flox) was 387 bp in length. The 549-bp fragment was produced by Cre-mediated recombination. Primers used are shown in Fig. 2A. B, detection of the Fhod3 protein by immunoblot analysis. Indicated amount of proteins prepared from the heart of TAM-treated Fhod3 iKO (Fhod3flox/−;MerCreMer+) and control littermate (Fhod3flox/−;MerCreMer−) mice were analyzed by immunoblot with anti-Fhod3. The loading amount was verified by myosin heavy chain bands stained with fast green. C, detection of the Fhod3 protein by immunoblot analysis. Proteins prepared from the heart of TAM-treated Fhod3 iKO (Fhod3flox/−;MerCreMer+) and TAM-treated control littermate (Fhod3flox/−;MerCreMer−) mice were analyzed by immunoblot with the three different anti-Fhod3 antibodies (anti-Fhod3-(650–802), anti-Fhod3-(873–974), and anti-Fhod3-C20) and anti-glyceraldehyde-3-phosphate dehydrogenase (GAPDH) antibody. D, survival curves of Fhod3 iKO (Fhod3flox/−;MerCreMer+, n = 72) and control littermate (Fhod3flox/−;MerCreMer−, n = 51; Fhod3flox/+;MerCreMer+, n = 42; Fhod3flox/+;MerCreMer−, n = 33) mice treated with TAM for the first 2 weeks. E, representative images of hearts of TAM-treated Fhod3 iKO (Fhod3flox/−;MerCreMer+) and TAM-treated control littermate (Fhod3flox/−;MerCreMer−) mice. These experiments have been repeated three times with similar results. Scale bars, 4 mm. F, histological analyses of hearts of TAM-treated Fhod3 iKO and TAM-treated control littermate mice. Longitudinal sections of hearts were stained with hematoxylin and eosin. Scale bars, 3 mm. G, heart-to-body weight and kidney-to-body weight ratios after TAM administration were determined. *, p < 0.05; N.S., not significant. H, quantitative real-time PCR analysis of Fhod1 gene expression in Fhod3 iKO and control littermate mice.