FIGURE

Fig. 5

ID

ZDB-FIG-210512-75

Publication

Zhao et al., 2021 - Heme-binding protein CYB5D1 is a radial spoke component required for coordinated ciliary beating

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Fig. 5

CrCYB5D1 localizes to the radial spoke stalk. Immunoblot analysis of equal amounts of protein from whole-cells, cell bodies, and isolated flagella of strains pf17 mC::CrCYB5D1 (A) and pf14 FLAG::CrCYB5D1 (B) probed with antibodies against mCherry and FLAG, respectively. (C) Representative immunogold electron micrograph of CrCYB5D1 localized at the stalk of the flagellar radial spoke using anti-mCherry antibody and a gold-conjugated secondary antibody. (Scale bar, 100 nm.) (D) The percentage of gold particles localized to different axoneme-associated components including the central pair (CP), radial spoke (RS) head, stalk and base, and the doublet microtubules (DMs). (E) Proteins from a whole-cell lysate of Crcyb5d1 mC::CrCYB5D1 were separated using sucrose density gradient centrifugation and collected in 24 fractions. Each fraction was subject to immunoblot analysis using antibodies against PF20, IC2, RSP1, RSP2, and mCherry. (F) Proteins from a flagellar membrane-matrix fraction of Crcyb5d1 mC::CrCYB5D1 were separated using sucrose density gradient centrifugation and collected in 20 fractions. Each fraction was subject to immunoblot analysis using antibodies against RSP1, RSP2, and mCherry.

Expression Data

Expression Detail

Antibody Labeling

Phenotype Data

Phenotype Detail

Acknowledgments

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