Sglt1 antibodies

Renal tissues were fixed in 10% formalin and embedded in paraffin. All histopathological scorings were made in the cortex using PAS-D stained renal sections and performed on coded slides. The percentage of tubular injury (criteria: epithelial flattening, tubular dilatation and brush border loss) and the degree of interstitial edema was estimated by a pathologist in a blinded fashion using a 4-point scale in ten randomly chosen, non-overlapping fields (200x magnification). Degree of injury was graded onto a scale from 0 to 4: 0 = normal; 1 = mild, involvement of less than 25% of the cortex; 2 = moderate, involvement of 25 to 50% of the cortex; 3 = severe, involvement of 50 to 75% of the cortex; 4 = extensive damage involving >75% of the cortex. Degree of edema was graded according: 0 = normal; 1 = mild edema; 2 = moderate; 3 = severe; 4 = extensive edema formation. For visualization of proximal tubular brush borders, renal sections were boiled in 10 mM Tris/1 mM EDTA (pH 9.0) and exposed to rabbit-anti-sodium/glucose transporter-1 (SGLT1) antibodies (Abcam), followed by power rabbit poly-HRP incubation and subsequently developed using 1% H₂O₂ and DAB (Sigma-Aldrich) in 0.05 M Tris-HCl (pH 7.9).

Chemicals were purchased from Sigma-Aldrich unless indicated otherwise. CFTR antibodies were obtained from the CF foundation. Antibodies against GRP78, IRE1α, XBP1s, and β-actin were purchased from Cell Signaling Technology. SGLT1 antibodies were purchased from Abcam and Invitrogen. The specificity of the SGLT1 antibody for IHC staining was confirmed by using an IgG isotype control, a secondary antibody–only control, and by using a nontargeting primary antibody control (Supplemental Figure 4). The secondary antibodies were from LI-COR Biosciences and Jackson ImmunoResearch Laboratories. The PAS staining kit was purchased from Thermo Fisher Scientific. The assay kits and antibody information are listed in Supplemental Table 4.