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. 2018 Jul 10;19(9):e45235. doi: 10.15252/embr.201745235

Figure EV1. Related to Fig 2: Bax is phosphorylated on residue S184.

Figure EV1

  1. Top two panels: Phosphorylation of Bax S184 was evaluated using lysates from MDA‐MB‐435 and MDA‐MB‐468 cells by Western blotting with the indicated antibodies (IB) after immunoprecipitation with the indicated antibodies (IP). Lower two panels: 5–10% of the total lysates (input) were probed for Bax and actin as expression and loading controls, respectively, by Western blotting with the indicated antibodies (IB).
  2. Top two panels: Phosphorylation of Bax S184 was evaluated using lysates from GFP‐Bax‐ and GFP‐Bax S184A‐expressing MDA‐MB‐468 cells by blotting with the antibodies indicated at the right after precipitation with the antibodies indicate below the panels. Lysates from untransfected cells (MDA‐MB‐468 WT) and IgG IP are used as negative controls. Lower panels: 5–10% of the total lysates were probed for GFP and actin as expression and loading controls, respectively.
  3. Localization of GFP‐Bax constructs in MDA‐MB‐468 cells. MDA‐MB‐468 cells transiently expressing GFP‐Bax S184A, GFP‐Bax, or GFP‐Bax S184E were lysed and separated into cytosolic (C), mitochondrial (M), or nuclear fractions (N) and then immunoblotted for GFP. GAPDH and CoxIV were immunoblotted for cytosolic and mitochondrial marker proteins, respectively.