Figure 7.

RAF1 oligomerization state in Rubisco mutants versus WT. (A) Immunoblot showing similar RAF1 content in rbcL or RBCS deletion mutants (ΔrbcL and ΔRBCS strains), and in WT, using antibodies directed against RAF1, Rubisco, and PsaD, as a loading control. Note that Rubisco accumulation was probed from a distinct membrane part obtained after the transfer of duplicated samples on the same gel. (B) Immunoblot of a 1D native PAGE of soluble extracts from ΔrbcL, ΔRBCS, and WT using RAF1 (left and middle panels) or Rubisco antibody (right panel), showing that RAF1 accumulates as an oligomer in the absence of LSU. RAF1-LSU complexes are indicated using the same symbols as in Figure 5. Note that the RAF1-LSU HMW complex found in the ΔRBCS is no longer detected in a WT background, whereas an additional low abundant RAF1 complex, indicated by a black star, is found. Red asterisks indicate antibody cross-reacting bands. (The left panel is a distinct experiment from the middle and right panels, which were separated on the same gel).