Figure 1

Olfr1393 is found in the renal proximal tubule.

(A) Renal segments were micro-dissected and reverse-transcribed with (RT-PCR) or without (mock RT-PCR) the reverse transcriptase enzyme. A representative gel from 1 of 3 separate mice is shown, and all bands were sequenced to confirm their identity. In all cases, Olfr1393 is found in all three regions of the proximal tubule (PT): S1, S2 and S3. (TAL, thick ascending limb (m, medullary, or c, cortical); DCT, distal convoluted tubule; CNT, connecting tubule; CCD, cortical collecting duct; OMCD, outer medulary collecting duct). For segments where sufficient amounts were collected, mock RT-PCR reactions were also performed. Olfr1393 was not detected in any of the mock reactions including all three regions of the proximal tubule. (B) RT-PCR for Olfr1393 was performed on mouse S3 cells (top) and BUMPT cells (bottom) to detect endogenous expression. Olfr1393 was detected in both cell lines in the RT-PCR (+) but not in the mock RT-PCR reactions (−). (C,D) Polarized MDCK cells were transiently transfected with flag-tagged Olfr1393. To detect Olfr1393, cells were stained with an anti-flag antibody either by (C) surface labeling the apical membrane in live, non-permeabilized cells or (D) staining for total Olfr1393 in fixed and permeabilized cells. In both cases, cells were also stained for gp135 (green) to mark the apical membrane. In (C) the merged image indicates colocalization between Olfr1393 and gp135 at the apical plasma membrane (Ap PM). All scale bars indicate 10 μM.