Interaction between endogenous PAQR3 and the components of VPS34 complex in HEK293T cells. The cell lysates were used for immunoprecipitation (IP) and immunoblotting (IB) with the antibodies as indicated.
Cytosol and Golgi fractionations were isolated from the livers of PAQR3‐deleted mice and their littermate controls. Equal protein amounts of cytosol and Golgi fractions were subjected to IB.
HEK293T cells were transiently transfected with Myc‐tagged PAQR3. At 24 h after transfection, the cells were treated with normal medium (NM), amino acid starvation (AS) or glucose starvation (GS), HBSS solution, or rapamycin (Rapa, 50 nM) for 4 h, respectively. The cell lysates were then used in IP and IB.
Four VPS34 complexes of MEFs were immunopurified using the indicated antibodies under NM, AS, or GS conditions. The relative abundance of VPS34‐binding partners was determined by IB. Each IP was normalized to the amount of VPS34.
HEK293T cells were transfected with the plasmids as indicated, and the cell lysates were used in IP and IB.
The lysates of WT or PAQR3 knockout MEFs were used in IP and IB.
MEFs were infected with control or PAQR3‐expressing lentivirus. Then, a quantitative immunodepletion assay was performed with increasing amounts of the indicated antibodies. Supernatants of MEF lysates after immunodepletion (Post‐IP) were examined to determine the level of VPS34 complex proteins.
