Figure 5. Ubiquitinated flagellar... | Rockefeller University Press

$Figure 5. Ubiquitinated flagellar proteins increase dramatically during flagellar resorption when IFT is blocked. (A) IFT temperature-sensitive mutants fla10, fla8, and fla3 were shifted to 32°C for 1 and 1.5 h, and immunoblots of the isolated, partially resorbed flagella were probed for ubiquitin (Ub) and IFT72/74. RSP3 was used as a loading control. WT, wild type. (B) HA∷ubiquitin was expressed in fla10 cells, and after the cells were incubated at 32°C for 60 min, resorbing flagella were isolated. Immunoblots of flagellar proteins were probed for ubiquitin and HA. (C) Isolated flagella (F), membrane/matrix (M), and axonemal fractions (Ax) of fla10 cells after shifting to 32°C for 1 h were analyzed on immunoblots probed with antibodies to ubiquitin, RSP3, and FMG-1. Most of the ubiquitinated proteins were in the membrane/matrix fraction, although some were retained by the axoneme. Molecular masses are given in kilodaltons.$

Figure 5.

Ubiquitinated flagellar proteins increase dramatically during flagellar resorption when IFT is blocked. (A) IFT temperature-sensitive mutants fla10, fla8, and fla3 were shifted to 32°C for 1 and 1.5 h, and immunoblots of the isolated, partially resorbed flagella were probed for ubiquitin (Ub) and IFT72/74. RSP3 was used as a loading control. WT, wild type. (B) HA∷ubiquitin was expressed in fla10 cells, and after the cells were incubated at 32°C for 60 min, resorbing flagella were isolated. Immunoblots of flagellar proteins were probed for ubiquitin and HA. (C) Isolated flagella (F), membrane/matrix (M), and axonemal fractions (Ax) of fla10 cells after shifting to 32°C for 1 h were analyzed on immunoblots probed with antibodies to ubiquitin, RSP3, and FMG-1. Most of the ubiquitinated proteins were in the membrane/matrix fraction, although some were retained by the axoneme. Molecular masses are given in kilodaltons.

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