Fig. 3.

A C-terminal di-leucine motif, ¹¹⁰⁵ERAQLL, confers LB targeting of ATP8A1. (A) Schematic representation of ATP8A1 and CDC50A protein topology [adapted from ATP8A2 in Coleman et al. (41) and refined by Hiraizumi et al. (42)] and strategy used for mutagenesis (LL = di-leucine; AA = di-alanine). (B) Representative confocal images of MLE15/WT cells expressing mCherry-ABCA3, and WT GFP-ATP8A1-WT or mutagenized GFP-ATP8A1 (AA1 ⁵³⁹ERYEAA; AA2 ¹¹⁰⁵ERAQAA; AA12 ⁵³⁹ERYEAA + ¹¹⁰⁵ERAQAA) as described in A. (Scale bar, 10 μm.) (C) Pearson’s correlation coefficients acquired from images of MLE15/WT cells described in B and D, and in SI Appendix, Fig. S3), comparing colocalization of GFP-ATP8A1-WT or GFP-ATP8A1-AA2 with organelle markers (n = 2 experiments; 15 to 20 cells; mean ± SE). (D) Representative confocal images of MLE15/WT cells expressing mCherry-ABCA3 and GFP-ATP8A1-AA2, and immunostained for RAB11A. (Scale bar, 10 μm.)