Rescue of VPS33B-KO imMKCLs with GFP-VPS33B shows the VPS33B-VPS16B complex localizes to recycling endosomes. (A) The endogenous VPS16B protein level in 2 VPS33B-KO imMKCL clones transfected with GFP-VPS33B or untransfected control was analyzed by immunoblotting (2 independent replicates per clone). (B) Live cell confocal fluorescence microscopy analysis of VPS33B-KO imMKCL cells cotransfected with GFP-VPS33B and Cherry-Rab11a plasmids. The main panels correspond to the first frame of a movie and display each channel separately and the merge. The region indicated with a dotted rectangle is shown below with frames taken every 5 seconds and highlighting an example of GFP-VPS33B puncta localization to a Cherry-Rab11a compartment over time.