Figure 4. BBS4 forms a complex with... | Rockefeller University Press

Figure 4.

$Figure 4. BBS4 forms a complex with other BBS proteins in the flagellum. (A) Proteins in the flagellar membrane-plus-matrix from BBS4HA21 (top) or wild type (bottom) were separated by sucrose gradient centrifugation, and fractions were analyzed by Western blotting using antibodies to the HA tag or BBS4 and IFT172, -139, and -46. White lines seperate images of the membranes used for this blot. (B) Silver-stained SDS gel of protein complexes affinity purified from the membrane-plus-matrix fraction of BBS4HA21 and wild-type (wt) flagella using anti-HA beads. Proteins identified by MS are indicated. (C) Western blot analysis of isolated flagella or whole-cell extracts from the strains indicated. The blots were probed with antibodies to BBS4 and, as loading controls, IC2 (flagella) or IFT139 (cells). ptx1-1 and ptx7-1 are control nonphototactic strains. (A–C) The positions of standard proteins and their molecular masses in kilodaltons are indicated.$

BBS4 forms a complex with other BBS proteins in the flagellum. (A) Proteins in the flagellar membrane-plus-matrix from BBS4HA21 (top) or wild type (bottom) were separated by sucrose gradient centrifugation, and fractions were analyzed by Western blotting using antibodies to the HA tag or BBS4 and IFT172, -139, and -46. White lines seperate images of the membranes used for this blot. (B) Silver-stained SDS gel of protein complexes affinity purified from the membrane-plus-matrix fraction of BBS4HA21 and wild-type (wt) flagella using anti-HA beads. Proteins identified by MS are indicated. (C) Western blot analysis of isolated flagella or whole-cell extracts from the strains indicated. The blots were probed with antibodies to BBS4 and, as loading controls, IC2 (flagella) or IFT139 (cells). ptx1-1 and ptx7-1 are control nonphototactic strains. (A–C) The positions of standard proteins and their molecular masses in kilodaltons are indicated.