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. 2015 Jul 31;16(9):1145–1163. doi: 10.15252/embr.201540759

Figure 7.

Figure 7

Loss of BNip3 induces increased dependence on autophagy for survival

  • A Effect of inhibiting glycolysis with 2-deoxyglucose for the growth rate of wild-type and BNip3 null MECs grown at 20% oxygen (performed in triplicate).
  • B Measurement of cell death measured by flow cytometric quantification of propidium iodide uptake in the presence or absence of 2-deoxyglucose.
  • C–H Immunohistochemical staining for LC3B (C–F) and p62/sqstm1 (G, H) on sections of wild-type (C, E, G; n = 4) and BNip3 null (D, F, H; n = 4) tumors at d80. Black scale bar is 50 μm. Red cut-out box in (C) and (D) is presented in (E) and (F), respectively.
  • I Western blot analysis of levels of BNip3, processed LC3B, p62/Sqstm1 and β-actin (loading control) in the presence or absence of 2-deoxyglucose or bafilomycin A1. Band intensity was calculated using ImageJ and standardized to the intensity of the loading control (β-actin). Fold change was determined relative to wild-type values in lane 1 for each protein measured.
  • J Measurement of cell death analyzed by flow cytometric quantification of propidium iodide uptake in the presence or absence of bafilomycin A1, hydroxychloroquine and/or 2-deoxyglucose, performed in duplicate experiments.

Data information: Results are expressed as the mean ± SEM. *P < 0.05, ***P < 0.001.