At least three lysates of EV1, EV2, #3 and #8 cells were subjected to western blot analysis. Representative blots and Coomassie-stained gels are shown (A, C, F); bar graphs indicate mean normalized intensity from all analyses +/- SEM (B, D, G). Statistical analysis was performed using one-way Anova. * indicates p<0.05 for both EV strains vs both PAM amiRNA strains. (A, B) Antibodies to CrPAM and modified forms of tubulin were used; acetylated and poly-glutamylated tubulin levels were lower in PAM amiRNA strains, but levels of α-tubulin were unchanged. (C, D) Antisera to clathrin heavy chain and Arf1 were used; PAM amiRNA strains had higher levels of clathrin heavy chain; Arf1 levels were not different. (E) Schematic of IFT and transition zone components in cilia; for the IFT motors, both Chlamydomonas and mammalian nomenclatures are shown. (F, G) Antisera to transition zone components CEP290 and NPHP4 were used; levels of both transition zone components were increased in PAM amiRNA mutant strains. (H) Maximum projection of optical sections of EV and PAM amiRNA C. reinhardtii cells labeled with antibodies to CEP290 (green) and acetylated tubulin (Ac tub, red). Merged images also show DAPI (blue), to locate the nucleus. Arrows point to CEP290 foci in the cell body. Enlargements at right show CEP290 localization at the base of cilia.