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Fig. 4 K48-linked ubiquitination of ATG16L1 is mediated by CX50. (A) HEK293 cells were transfected with HA-ATG16L1 or Myc-Ub with or without Flag-CX50. After 24 h transfection, IP was performed with HA affinity gels on cells with or without MG132 treatment for 6 h, and ubiquitination was analyzed using anti-Myc. WCL: whole cell lysis. (B) Representative images showing the distribution of ATG16L1 and Ub in lenses from 72 hpf WT and cx50-deficient zebrafish. Scale bar: 10 µm. (C-F) The HEK293 cells were transfected with indicated plasmids for 24 h, and the IP using HA affinity gels was subjected to immunoblotting using the indicated antibodies.