P22phox fl 195

Primary antibodies used were NOX1 [21 (link)]; NOX2 (53/gp91^phox, BD); V5-tag (V5-10), Vinculin (hVIN-1), β-Tubulin (ONS.1A6), F-Actin (polyclonal) (all Sigma-Aldrich); HA-tag (16B12, Covance); p22^phox (FL-195, Santa Cruz); p47^phox and p67^phox [22 (link)].

Nuclear and cytoplasmic protein was extracted and fractionated as previously described [9 (link)]. Western blotting was performed using the following antibodies: Ref-1 (E-17, 1:250, Cat sc-9919, and C-4, 1:250, Cat sc-17774, Sana Cruz), Acetylated Ref-1 (1:1000, Cat D12045, Ancell), p22^phox (FL-195, 1:200, Cat sc-20781, Santa Cruz), Beclin-1 (D40C5, 1:1000, Cat #3495, Cell Signaling), LC3B ((D11)XPTM, 1:1000, Cat #3836, Cell Signaling), p62 (D5E2, 1:1000, Cat #8025, Cell Signaling), EGR1 (15F7, 1:250, Cat #4153, Cell Signaling), PTEN (138G6, 1:500, Cat #9559, Cell Signaling), Akt (pan; C67E7, 1:1000, Cat #9916, Cell Signaling), phospho-Akt (ser473, Cat #9916, 1:1000, Cell Signaling), RAC1 (1:200, Santa Cruz), and β-actin (1:5000, Santa Cruz) as described previously [6 (link)]. Samples from A549, control KD, p22^phox KD, and Ref-1 KD cells were also used for immunoprecipitation, which was performed as described previously [48 (link)].