Figure 1.

Gp91phox and p22phox segregate to the buoyant low-density detergent (Brij-58, Brij-98) resistant membrane fraction. Ra2 microglia (A–H) or HL60 (I–Q) cells were lysed in Brij-58 (A–D, I–M) or Brij-98 (E–H, N–Q), in some cases with prior mβCD-facilitated cholesterol extraction (B, F, J), and the lysate was centrifuged to equilibrium in a 35–15% sucrose gradient. Recovered fractions (no. 12 is the bottom, high-density fraction of the gradient) and the solubilized pellet (P) were analyzed by Western blotting using antibodies against gp91phox (gp91), p22phox (p22), transferrin receptor (TfnR), β1-integrin (β1), flotillin-2 (Flo-2), or Lyn as indicated.