Picture from Arata, Yukinobu et al. (2020) MicroPubl Biol
Figure 1: Biofilm formation, the establishment of OP50-derived fliC and fimH knockout strains, and bacterial motility assay. Biofilm formation on glass (A) and polystyrene (B). Biofilm formed by indicated E. coli strains on glass and polystyrene quantified by CV staining. Optical absorbance at 600 nm was measured in three independent experiments and compared with paired student t-tests; *p < 0.05, ** p < 0.005. Biofilm formation ability of OP50 fliC::Km, OP50 fimH::Km, and BL21 should be studied further with rigorous experiments and statistical testing. (C and D) Isolation of OP50 fimH::Km and OP50 fliC::Km. (C) Schematic structure of fliC and fimH knockout alleles with position and direction of primers for PCR. Km cassette was integrated in gene locus to form fimH::Km and fliC::Km. (D) Confirmation of the presence of fimH::Km and fliC::Km in OP50-derived E. coli colonies via P1 transduction. Bacteria obtained from Km-resistant colonies were subjected to PCR with the indicated primers. DNA fragments of expected sizes from the genomic structure of the fliC::Km and fimH::Km knockout alleles were amplified with a primer set with K1 but not K2 primer. (E) Motility of E. coli strains. Motility of indicated E. coli strains was compared by colony size after cultivation on soft agar LB plates.
Picture from Browning H et al. (1996) Development "A sperm-supplied factor required for embryogenesis in C. elegans."
Figure 6. Immunofluorescence analyses of SPE-11 localization during spermatogenesis. In each confocal micrograph (A,C,D) spermatogenesis proceeds from left to right. SPE-11 is stained with affinity purified antiserum to SPE-11, DNA is stained with propidium iodide or 4, 6 diamidino-2-phenylindole (DAPI), andnuclear envelopes are stained with a monoclonal antibody (mAb414) to nuclear pore complex (NPC) proteins (Davis and Blobel, 1986). Scale bar, 10 µm. (A) Confocal images of a wild-type male gonad double labeled with antiserum to SPE-11 and propidium iodide. Each micrograph is the projection of a Z series containing seven sequential focal planes including approximately half the gonad thickness. SPE-11 first appears as dots in the nuclei of primary spermatocytes. The dots appear to coalesce before the first meiotic division. During the first and second meiotic divisions, SPE-11 is diffuse throughout the cells (bracket). SPE-11 is localized in a ring around the nuclei of sperm. (B) Conventional epifluorescence micrographs of sperm double labeled with antiserum to SPE-11 and DAPI. SPE-11 is perinuclear in sperm. Inset: Four spermatids attached to a single residual body. Some staining is detectable in the residual body. (C) Confocal images of primary spermatocytes double labeled with antiserum to SPE-11 and propidium iodide. The micrographs are projections of a Z series that includes the entire depth of the tissue. As the DNA becomes condensed in preparation for the first meiotic division, SPE-11 forms a fenestrated ring around the DNA. As discrete chromosomes become visible, SPE-11 relocalizes throughout the nucleus and is sometimes localized between the chromosomes. (This is also evident from analysis of individual focal planes; data not shown.) (D) Confocal images of primary spermatocytes double labeled with antiserum to SPE-11 and NPC antibodies. The micrographs are projections of a Z series that include the entire depth of the tissue. A merge of the NPC and SPE-11 projections demonstrates that the dynamic relocalization that SPE-11 undergoes in primary spermatocytes occurs within the nucleus. Spermatogenesis proceeds from left to right, with the exception of a few sperm (small rings stained with antiserum to SPE-11) in the upper left portion of the micrographs. There is no detectable staining of these sperm with the NPC antibody.