Figure 6.
Generation of PEPCK-Cre-S1P1 floxed mice results in reduced copy number of S1P1Rs in kidney cortex. (A) Genotyping of PEPCK-Cre and S1P1 floxed mice. Agarose gel analysis of PCR products amplified from tail DNA. Lane 1, molecular weight markers; lane 2, PEPCK-Cre; lane 3, S1P1fl/wt; lane 4, PEPCK-CreS1P1fl/wt; lane 5, S1P1fl/fl; lane 6, PEPCK-CreS1P1fl/fl. Size of predicted PCR products (number of bp) is shown at right side of gel. (B and C) Immunofluorescent localization of YFP reporter (GFP immunoreactivity, green) indicative of Cre recombinase expression in kidney (B) and liver (C) sections of PEPCK-CreRosa26YFP/wt mice. Nuclei were labeled with DAPI, blue. Bar = 40 μm. (D) Representative gel of S1P1R mRNA levels in different tissues from PEPCK-Cre, PEPCK-CreS1P1fl/wt, and PEPCK-CreS1P1fl/fl mice (lanes from left to right). Regions of cortex (CTX) and inner medulla/papilla (IM) were dissected from kidney. (E) Expression of S1P1R (mRNA levels relative to GAPDH, determined by real-time RT-PCR) in kidney cortex (Kid-CTX) and medulla (Kid-IM), liver, and lung from PEPCK-Cre, PEPCK-CreS1P1fl/wt, and PEPCK-CreS1P1fl/fl mice.