(A) Western blots of human eIF3 subunits in crude lysates (L) or immunoprecipitate (IP) with eIF3b primary antibody from either WT or EIF3H KO (ΔeIF3h) HEK293T cells.
(B) Schematics depicting N-terminally FLAG-tagged eIF3d or e constructs used for FLAG IPs in (C–F). PCI domains are represented with blue rectangles and FLAG tags are black rectangles. The amino acids included in truncated constructs are also shown.
(C–F) Western blots of anti-FLAG immunoprecipitations using transiently transfected FLAG-tagged constructs, as indicated. The cell lines, WT or EIF3H KO (ΔeIF3h), used for expression are indicated over each pair of western blots. The probed eIF3 subunit is indicated to the left or right of each blot. Subunits marked with an asterisk indicate the bands of the FLAG-tagged subunit. (C–F) Contain samples from crude L, unbound by anti-FLAG antibody flow through (FT), wash (W), and FLAG IP.