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Fig. 2 CrCYB5D1 is essential for normal motility in Chlamydomonas. (A) Schematic diagram of the genomic structure of CrCYB5D1 illustrating the location of the insertion in the Crcyb5d1 mutant. The AphVIII expression cassette is inserted into the fourth exon. (B) PCR analyses of CrCYB5D1 using genomic DNA of HS211 and Crcyb5d1 as templates, using the primers shown in A. (C) Transcriptional analysis of CrCYB5D1 in the indicated strains using RT-PCR. (D) Immunoblot of whole-cell lysates from the indicated strains using antibody against mCherry; α-tubulin served as a loading control. (E) The swimming tracks of individual cells from the indicated strains were recorded for a 5 s period using a micro image analyzer (Olympus). The red and green lines indicate the straight and curved motion tracks, respectively; dark blue dots are nonmotile cells. (Scale bars, 100 μm.) (F) Bar graph showing the percentage of forward swimming cells in the indicated strains. (G) The forward swimming velocity of cells in the indicated strains. For F and G the number of cells (n) measured is indicated. ***P < 0.001.