Figure S5.
AMPK inhibits autophagosome maturation. (A) Western blots probed for ULK1 phosphorylated at S556 of control U2OS cells, and U2OS cells expressing Halo-ATG13, Halo-ATG2, Halo-ULK1, or Halo-WIPI2 grown in complete media, or media lacking glucose. (B) Western blots probed for ULK1 phosphorylated at S556 of control U2OS cells, and U2OS cells expressing Halo-ATG13, or Halo-WIPI2 grown in complete media, or media lacking glucose in the absence or presence of 1 µM or 20 µM of the AMPK inhibitor BAY3827. Total protein staining was carried out prior to protein transfer onto a nitrocellulose membrane and the membrane was cut into two pieces for western blot procedures. Both membrane pieces were stained in the same dish and exposed simultaneously. (C) Western blots probed for ULK1 phosphorylated at S556 of control U2OS cells, and U2OS cells expressing Halo-ATG13, or Halo-WIPI2 grown in complete media, in the absence and presence of 1 µM or 20 µM of the AMPK activator MK8722. (D) Live-cell images of ULK1-Halo, Halo-ATG13, or Halo-ATG2A and GFP-LC3B in the indicated media conditions in the presence and absence of the AMPK activator MK8722 or the AMPK inhibitor BAY3827 (scale bar = 5 µm). Source data are available for this figure: SourceData FS5.

AMPK inhibits autophagosome maturation. (A) Western blots probed for ULK1 phosphorylated at S556 of control U2OS cells, and U2OS cells expressing Halo-ATG13, Halo-ATG2, Halo-ULK1, or Halo-WIPI2 grown in complete media, or media lacking glucose. (B) Western blots probed for ULK1 phosphorylated at S556 of control U2OS cells, and U2OS cells expressing Halo-ATG13, or Halo-WIPI2 grown in complete media, or media lacking glucose in the absence or presence of 1 µM or 20 µM of the AMPK inhibitor BAY3827. Total protein staining was carried out prior to protein transfer onto a nitrocellulose membrane and the membrane was cut into two pieces for western blot procedures. Both membrane pieces were stained in the same dish and exposed simultaneously. (C) Western blots probed for ULK1 phosphorylated at S556 of control U2OS cells, and U2OS cells expressing Halo-ATG13, or Halo-WIPI2 grown in complete media, in the absence and presence of 1 µM or 20 µM of the AMPK activator MK8722. (D) Live-cell images of ULK1-Halo, Halo-ATG13, or Halo-ATG2A and GFP-LC3B in the indicated media conditions in the presence and absence of the AMPK activator MK8722 or the AMPK inhibitor BAY3827 (scale bar = 5 µm). Source data are available for this figure: SourceData FS5.

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