(A) MDCK cells were grown on Transwells for 4-5 days post confluence, fixed with paraformaldehyde, extracted with methanol, and stained for endogenous flotillin-1. (B) As in A except cells were stained for endogenous flotillin-2. (C) Stable MDCK cell lines expressing GFP-Rab11-FIP2 wild type (WT), a N-terminal truncation minus the C2 domain (ΔC2) or a double point mutation (SARG) were grown on Transwells for 4-5 days post confluence, fixed with paraformaldehyde, extracted with methanol, and stained for endogenous flotillin-1 (second column, red in merge). The GFP-FIP2 constructs are shown in the first column (green in merge). Black arrowheads indicate where the XY and XZ slices were taken. (D) As in C, except cells were stained for endogenous flotillin-2. Bar=5 μm.