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. 2017 Nov 1;8(62):105440–105457. doi: 10.18632/oncotarget.22241

Figure 4. NOX4 67 kDa- and NOX4D 28 kDa–generated pro-survival ROS require AKT activation.

Figure 4

(A) Western blot analysis of AKT signalling in FLT3-ITD expressing MV4-11 cells following treatment with LY294002 (20 μM, 30 μM and 50 μM) for 16 h. β-actin and total protein were used as loading controls. (B) NOX4 67 kDa, NOX4D 28 kDa and p22phox protein expression in membrane and soluble nuclear fractions of MV4-11 cells following treatment with LY294002 for 16 h at indicated concentrations. Equal loading of samples is shown by total protein and verification of subcellular fractions were assessed by probing for nuclear-localised NUP98 and membrane-localised calreticulin. Western blot analysis is representative of three independent experiments. (C) (i) Flow cytometric analysis of mean relative PO1 fluorescence in MV4-11 cells treated with LY294002 for 16 h at indicated concentrations. (ii) Bar chart shows relative mean PO1 fluorescence of treated cells expressed as % of control. Results are representative of three independent experiments. Asterisks indicate statistically significant differences (****p<0.0001) as analysed by Student's t-test. Error bars are representative of SD.