Figure 2.

ABCG1-GFP alters cholesterol detergent solubility. (A) Control and ABCG1 cells were incubated with PBS or SMase, washed, fixed and immunostained with lysenin as described in “Experimental Section”. (B) Control and ABCG1 cells were incubated with cold PBS or 1% TX-100 detergent for 30 min, washed, fixed and immunostained with lysenin as described in “Experimental Section”. (C) Control and ABCG1 cells pre-treated with or without SMase, were extracted with 1% TX-100 on ice for 30 min, fixed, filipin stained, and imaged by confocal microscopy as described in “Experimental Section.” (D) Higher magnification of filipin-stained AGCG1 cells clearly reveals that LE FC (a) is removed by TX-100 (b). Enlargements of the boxed regions in (a) and (b), shown in (c) and (d), respectively, reveal that TX-100 treatment depleted FC from PM domains (arrowheads).