Figure 2. Loss of ATP8B1 in polarized... | Rockefeller University Press

Figure 2.

$Figure 2. Loss of ATP8B1 in polarized Ls174T:W4 cells results in the formation of an enlarged apical membrane and causes loss of singularity. (A) Polarized Ls174T:W4 cells infected with control short hairpin or shATP8B1 stained with phalloidin to visualize the actin cytoskeleton. Blue outlining indicates apical membrane, and yellow outlining indicates basolateral membrane. (B) Apical membrane size quantified by determining the fraction of the plasma membrane bearing microvilli as visualized by actin staining. *, P < 0.00002. (C) Western blot of W4 cell and ATP8B1-depleted W4 cell lysates probed for ATP8B1 and Na/K ATPase. (D) Localization of the apical recycling endosomal marker dsRed-Rab11 in control- and ATP8B1-depleted W4 cells. (E) Quantification of cells with multiple brush borders in polarized control or ATP8B1-depleted W4 cells. Numbers in parentheses indicate total number of cells with multiple apical membranes in three independent experiments. Error bars are SEM from three experiments. *, P < 0.0003. (F) Example of ATP8B1-depleted cell that has formed two discrete brush border patches. Bars, 5 µm.$

Loss of ATP8B1 in polarized Ls174T:W4 cells results in the formation of an enlarged apical membrane and causes loss of singularity. (A) Polarized Ls174T:W4 cells infected with control short hairpin or shATP8B1 stained with phalloidin to visualize the actin cytoskeleton. Blue outlining indicates apical membrane, and yellow outlining indicates basolateral membrane. (B) Apical membrane size quantified by determining the fraction of the plasma membrane bearing microvilli as visualized by actin staining. *, P < 0.00002. (C) Western blot of W4 cell and ATP8B1-depleted W4 cell lysates probed for ATP8B1 and Na/K ATPase. (D) Localization of the apical recycling endosomal marker dsRed-Rab11 in control- and ATP8B1-depleted W4 cells. (E) Quantification of cells with multiple brush borders in polarized control or ATP8B1-depleted W4 cells. Numbers in parentheses indicate total number of cells with multiple apical membranes in three independent experiments. Error bars are SEM from three experiments. *, P < 0.0003. (F) Example of ATP8B1-depleted cell that has formed two discrete brush border patches. Bars, 5 µm.

This Feature Is Available To Subscribers Only