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Results: 4

1.
Figure 4

Figure 4. From: Profilin I is essential for cell survival and cell division in early mouse development.

Levels of profilin I in platelets and thymocytes from heterozygous mutant mice. (a) Equal amounts of platelet lysate from pfn1wt/wt and pfn1ko/wt mice were separated by SDS/PAGE, transferred, and probed with a profilin I-specific antibody (Bottom), or the gel was stained with Coomassie blue (Top). The amount of profilin I is reduced by about 50% in the heterozygotes. (b) Equal amounts of thymocyte lysate from pfn1wt/wt and pfn1ko/wt mice were subjected to SDS/PAGE and probed with a profilin I-specific antibody. The filter was reprobed with a (capG)-specific antibody to normalize for loaded protein amounts. Note the reduction of profilin I in the mutant thymocytes.

Walter Witke, et al. Proc Natl Acad Sci U S A. 2001 March 27;98(7):3832-3836.
2.
Figure 1

Figure 1. From: Profilin I is essential for cell survival and cell division in early mouse development.

Expression of profilin I and profilin IIA protein in mouse tissues and during development. (a) Equal amounts of tissue extracts from adult mice were separated by SDS/PAGE, transferred, and probed with affinity-purified polyclonal antibodies raised against profilin I (P I) and profilin II (P II). (b) Lysates from ES cells and embryos at different stages [embryonic days 10–20 (E10–E20)] were tested for levels of profilins I and IIA. (c) Absolute amounts of profilins I and IIA expressed in various mouse tissues in percentage of total cellular protein.

Walter Witke, et al. Proc Natl Acad Sci U S A. 2001 March 27;98(7):3832-3836.
3.
Figure 3

Figure 3. From: Profilin I is essential for cell survival and cell division in early mouse development.

In vitro development of two-cell embryos from wild-type (pfn1wt/wt) and heterozygous (pfn1ko/wt) crossings. Embryos were flushed at E1.5 and cultured for 2 days according to standard procedures (34) to allow development into morulas/blastocysts. (a) Embryos from a pfn1wt/wt mating after 3.5 days. (b) Embryos from a pfn1ko/wt mating after 3.5 days. Note that a significant number of embryos from the mutant cross are growth-arrested and deteriorated. (c) Staining of a wild-type morula for profilin I (green), F-actin (red), and DNA (blue). Embryos were stained with affinity-purified anti-profilin I antibody and an FITC-labeled secondary antibody, tetramethylrhodamine B isothiocyanate-phalloidin for F-actin, and 4′,6-diamidino-2-phenylindole for DNA. Details of the immunostaining protocol are available upon request.

Walter Witke, et al. Proc Natl Acad Sci U S A. 2001 March 27;98(7):3832-3836.
4.
Figure 2

Figure 2. From: Profilin I is essential for cell survival and cell division in early mouse development.

Generation of profilin I knockout mice. (a) Map of the mouse profilin I gene (Top), the targeting vector (Middle) consisting of an 8.5-kb BglII (Bg) fragment, and the targeted locus (Bottom) are shown. Filled rectangles indicate coding exons 1, 2, and 3. The arrows (Bottom) indicate the primers used to identify the targeted alleles for the PCR analysis of blastocyst DNA. (b) Homologous recombination in ES cells is indicated by generation of an 11.8-kbp fragment (marked with an asterisk) caused by insertion of the neo gene into the wild-type allele of 10 kbp. DNA was tested with the indicated probes (lanes 1 and 2, profilin probe; lane 3, neo probe) after restriction digestion with HindIII (H). (c) Analysis of DNA from litters of a heterozygous breeding. DNA was digested with BamHI and probed with the indicated probe. The targeted allele (5.8 kbp) is marked with an asterisk.

Walter Witke, et al. Proc Natl Acad Sci U S A. 2001 March 27;98(7):3832-3836.

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