Fig. 8

miR-320a regulates the ubiquitination of AMPK by targeting CBL. A A549 cells were treated with the NC mimic and miR-320a mimic, and then cells were collected for Western blotting to show the expressions of CBL and AMPK. B A549 cells were treated with NC mimic or miR-320a mimic for 24 h, followed by MG132 treatment (20 μM) for 2 h. Expression of AMPK was assessed by Western blotting analysis. C The endogenous interaction of CBL and AMPK was tested in A549 cells. D Flag-CBL and Myc-AMPK plasmids were transfected into HEK-293 T cells, and CO-IP detected the interaction between CBL and AMPK. E Ubiquitination of AMPK was analyzed in WT and si-CBL A549 cells. F Ubiquitination of AMPK was analyzed in A549 cells transfected with NC mimics, and miR-320a mimics. G Flag-CBL, Myc-AMPK, and HA-ub plasmids were transfected alone or cotransfected into HEK-293 T cells, and the ubiquitination of AMPK was detected by CO-IP. H HEK-293 T cells were transfected with ubiquitin mutants (K48R and K63R), Myc-AMPK, and Flag-CBL. interaction between AMPK and ubiquitin was detected by Co-IP. I AMPK and p-JNK were detected by Western blotting. Blots were representative of six independent experiments