Gb112093

Immunohistochemistry for the protein expression of mouse kidneys was assessed using the streptavidin–peroxidase immunohistochemical method. For immunohistochemistry staining, deparaffinized sections were boiled in citrate buffer for antigen retrieval. Then, the sections were permeabilized in 1× PBS containing Triton X-100 (0.1%) for 10 min and incubated with anti-α smooth muscle actin (α-SMA, GB11044, Servicebio, Wuhan, China), antifibronectin (GB112093, Servicebio), anticollagen I (PB0981, BOSTER, Wuhan, China), anti-PKM2 antibody (4053S, CST, Boston, United States), anti-p-PKM2 (PA5-105498, Invitrogen, California, United States), anti-CD206 (ab125028, Abcam, Cambridge, United Kingdom), anti-inducible nitric oxide synthase (iNOS, ab178945, Abcam) at 4°C overnight. Immunohistochemical staining was performed using an UltraSensitive S-P kit (kit 9706, Maixin-Bio, Jinan, China). The selected regions were captured under a light microscope.

Immunofluorescence analysis was performed with mouse antibodies of F4/80 (Servicebio, GB11027), CD44 (Servicebio, GB112054), FN1 (Servicebio, GB112093) and SPP1 (Servicebio, GB11500). The tumor tissue was firstly fixed with paraformaldehyde, and embedded in paraffin, and then used for immunofluorescence staining.