Anti lotus tetragonolobus lectin fl 1321

Kidney cryosections were fixed with 4% paraformaldehyde solution for 15 min at room temperature. Primary antibodies were as follows: anti‐active‐β‐catenin (4270s; Cell Signaling Technology), anti‐TOMM20 (ab186735; Abcam), anti‐Lotus Tetragonolobus Lectin (LTL) (FL‐1321; VECTOR Laboratories), anti‐Peanut Agglutinin (PNA)(FL‐1071; VECTOR Laboratories), anti‐Dolichos Biflorus Agglutinin (DBA)(FL1031; VECTOR Laboratories). After washing with TBS‐T, slides were incubated with Cy2 or Cy3‐conjugated donkey anti‐mouse or anti‐rabbit IgG (Jackson Immuno‐Research Laboratories, West Grove, PA). Nuclei were stained with DAPI (C1006, Beyotime) according to the manufacturer's instructions. Images were taken by confocal microscopy (Leica TCS SP2 AOBS, Leica Microsystems, Buffalo Grove, IL).

Immunofluorescence staining was performed with routine protocol (Zhou et al., 2018 (link)). Frozen kidney sections or the cover slides of cell culture were fixed with 4% paraformaldehyde at room temperature for 15 min. Primary antibodies were incubated after blocking with 10% donkey serum at room temperature for 1 h. All primary antibodies used were as follows: anti-fibronectin (F3648; Sigma-Aldrich), anti-TOMM20 (ab186735; Abcam), anti-γ-H2AX (ab26350; Abcam), anti-CXCR2 (Ab61100; Abcam), anti-β-catenin (610154; BD Transduction Laboratories), anti-Lotus Tetragonolobus Lectin (LTL) (FL-1321; VECTOR Laboratories), anti-Peanut Agglutinin (PNA) (FL-1071; VECTOR Laboratories), anti-Dolichos Biflorus Agglutinin (DBA) (FL1031; VECTOR Laboratories). Then the slides were treated with Cy3-or Cy2-conjugated secondary antibodies (Jackson Immuno-Research Laboratories, West Grove, PA, United States) for 1 h at room temperature. Nuclei were stained using DAPI (Sigma-Aldrich) under the manufacturer’s specifications.