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. 2016 May 3;18(11):1596–1610. doi: 10.1111/cmi.12599

Figure 4.

Figure 4

Immunoprecipitation of mutant PfSec22 and PfARO proteins using antibodies against the c‐Myc tag. Human embryonic kidney 293E cells were co‐transfected with plasmids coding for the expression of mutant c‐Myc‐tagged PfSec22 or PfARO (which had the predicted palmitoylated cysteine residues mutated to alanine residues), along with FLAG‐tagged PfDHHC5 or the control vector (CD4). The mutant PfSec22 and PfARO proteins were immunoprecipitated from cell lysates using α‐c‐Myc antibody. The proteins were separated by SDS‐PAGE and visualized by immunoblot, using α‐c‐Myc antibody from a different species. A. Immunoprecipitation of PfSec22 point mutants (PfSec22‐C2dA, PfSec22‐C8dA and PfSec22‐C2C8dA) along with wild‐type PfSec22. The amino acid sequence of the N‐terminal region of PfSec22 is also shown with the cysteine residues of interest highlighted in red. B. Immunoprecipitation of PfARO point mutants (PfARO‐C5dA, PfARO‐C6dA and PfARO‐C5C6dA) along with wild‐type PfARO. The amino acid sequence of the N‐terminal region of PfARO is also shown with the cysteine residues of interest highlighted in red.