(A) In vivo ubiquitination assay in HEK293T cells transfected with the indicated plasmids was followed by IB.
(B) LKB1-deficient HeLa cells with stable restoration of the indicated Flag-LKB1 were subjected to IB.
(C) Control- or LKB1-knockdown Hep3B cells with stable transduction of the indicated shRNA non-targeting myc-LKB1 cultured in the absence of glucose for the indicated time points were subjected to IB.
(D) IPs of exogenous Flag-LKB1 by anti-Flag antibody from HEK293T cells transfected with the indicated Flag-LKB1 were subjected to in vitro LKB1 kinase assay followed by IB.
(E) Hep3B cells with stable transduction of vector (Vec) or Skp2 along with the indicated Flag-LKB1 cultured in the presence (+) or absence (−) of glucose were subjected to IB.
S.E., short exposure; L.E., long exposure.
See also Figure S3.