*4.6. Immunohistochemical Staining*

Renal lymphatic vessels were collected from rats, fixed in the 4% paraformaldehyde, and embedded in paraffin. Three-micron paraffin sections were stained with the standard

protocol. For NKCC1 staining, tissues were deparaffinized and then antigen retrieved with citrate buffer (pH = 6.0). After blocking with 2.5% normal horse serum, the tissue was incubated with anti-NKCC1 antibody (Boster Bio, 1:1000, Pleasanton, CA, USA) overnight and then incubated with anti-rabbit secondary antibody. Negative control was prepared by omitting the primary antibody.
