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. 2023 Apr 14;299(6):104718. doi: 10.1016/j.jbc.2023.104718

Figure 2.

Figure 2

The VPS33B–VPS16B complex contains multiple copies of each component.A–D, SDS-PAGE immunoblots show differentially tagged VPS33B and VPS16B expressed in transfected HEK293 cells can co-IP a second copy of each protein. A, probing for FLAG, HA, Myc, and GAPDH (bottom) of lysates from HEK293 cells cotransfected with HA-VPS33B + Myc-VPS16B, and with/without VPS16B-FLAG as indicated. B, in lysates from triple-transfected cells, anti-FLAG antibody co-IP’s HA-tagged VPS33B and Myc-tagged VPS16B, confirming at least two copies of VPS16B in the complex. C, probing of lysates from cells cotransfected with HA-VPS33B + Myc-VPS16B, and with/without VPS33B-FLAG. D, in lysates from triple-transfected cells, anti-FLAG antibody co-IP’s HA-tagged VPS33B and Myc-tagged VPS16B, confirming at least two copies of VPS33B in the complex. co-IP, coimmunoprecipitation; HA, hemagglutinin; HEK293, human embryonic kidney 293 cell line; VPS, vacuolar protein sorting protein.