FACS Strategy for Adult Duct Cells
The sequence of the sorting work flow is shown from left to right in (A) to (E). Mouse liver non-parenchymal (NPC) cells were labeled with MIC1-1C3, ST14, CD26, CD31, CD45, and CD11b.
(A and B) Cells were sequentially gated based on cell size (forward scatter [FSC] versus side scatter [SSC]) (A) and singlets (FSC versus trigger pulse width) (B).
(C) Dead cells and debris were excluded by detection of propidium iodide (PI) positivity. Concurrently a combination of CD45, CD31, and CD11b antibodies was used for depleting blood, endothelium, and Kupffer cells.
(D) CD26 (DPPIV) was used for hepatocyte staining.
(E) MIC1-1C3+ cells can be subdivided into two populations: ST14 high (ST14hiM+) and ST14 low (ST14loM+).
(F) Size and scatter properties of fully gated ST14hiM+ cells.
n = 10 independent mice. See also Figure S1.