Figure 1. Flotillin microdomains are... | Rockefeller University Press

Figure 1.

$Figure 1. Flotillin microdomains are absent in flotillin 1−/− mice. (a) Gene targeting strategy. Positions of primers used for PCR genotyping are shown as black arrows; the primers are labeled a, b, c. (b) Genotyping of wild-type, C57Bl6/J flotillin 1+/+ (+/+), flotillin 1 knockout C57Bl6/J flotillin 1−/− (−/−), and flotillin 1+/− heterozygous mice (+/−). (c) Flotillin 1−/− mice lack flotillin 1, and flotillin 2 expression in these mice is drastically reduced. Western blots using antibodies against flotillin 1 and flotillin 2. MEF, mouse embryonic fibroblast. (d) Residual flotillin 2 in flotillin 1−/− MEFs is not concentrated in flotillin microdomains. Staining of mixed culture of flotillin 1−/− and wild-type MEFs with flotillin 1 and flotillin 2 antibodies. Bar, 20 µm. (e) Residual flotillin 2 in flotillin 1−/− MEFs is not present in detergent-resistant membranes. Detergent-resistant membranes (fractions 9 and 10, boxed) were floated in a sucrose density gradient. Western blots of samples from control (+/+) and flotillin 1−/− (−/−) mice were probed with antibodies to flotillin 2.$

Flotillin microdomains are absent in flotillin 1^−/− mice. (a) Gene targeting strategy. Positions of primers used for PCR genotyping are shown as black arrows; the primers are labeled a, b, c. (b) Genotyping of wild-type, C57Bl6/J flotillin 1^+/+ (+/+), flotillin 1 knockout C57Bl6/J flotillin 1^−/− (−/−), and flotillin 1^+/− heterozygous mice (+/−). (c) Flotillin 1^−/− mice lack flotillin 1, and flotillin 2 expression in these mice is drastically reduced. Western blots using antibodies against flotillin 1 and flotillin 2. MEF, mouse embryonic fibroblast. (d) Residual flotillin 2 in flotillin 1^−/− MEFs is not concentrated in flotillin microdomains. Staining of mixed culture of flotillin 1^−/− and wild-type MEFs with flotillin 1 and flotillin 2 antibodies. Bar, 20 µm. (e) Residual flotillin 2 in flotillin 1^−/− MEFs is not present in detergent-resistant membranes. Detergent-resistant membranes (fractions 9 and 10, boxed) were floated in a sucrose density gradient. Western blots of samples from control (+/+) and flotillin 1^−/− (−/−) mice were probed with antibodies to flotillin 2.