Fig. 2.
BAP31, VAPA, and ESYT1 are interaction partners of ICP47SBP-arrested PLCs in moDCs. Monocytes (mono), imDCs, mDCs, and Raji cells were solubilized in 1% (w/v) digitonin and affinity purified in the presence (+) or absence (−) of ICP47SBP. Proteins were separated on SDS-PAGE and subsequently immunoblotted. To avoid donor dependencies, two to three donors were pooled for each primary cell type. The left panel shows cell lysate material after solubilization and ultracentrifugation (solubilizate). On the right panel, the eluates after pulldown are presented. Classical components of the PLC (A) TAP1, ERp57, and MHC I and (B) TAP2, calreticulin (Crt), and tapasin (Tsn) are detected in the ICP47SBP pull-down. (C) ESYT1, VAPA, and BAP31 are detected by immunoblotting. GAPDH served as a loading control.