Anti pfkfb3 antibody

Total RNA was extracted using TRIzol (Takara) and cDNA of each group was synthesized with the PrimeScript™ RT reagent kit (Takara). qPCR was performed with the Roche LightCycler 480II real-time PCR detection system (Roche, Basel, Switzerland). Expression level of each gene was normalized to that of β-actin. The primers for qRT-PCR are listed in Supplementary Table 2.
Western blotting (WB) was performed as described previously (17 (link)). The antibodies used were as follows: anti-AHNAK antibody (sc-390743, Santa Cruz Biotechnology), anti-NFATC1 antibody (sc-7294, Santa Cruz Biotechnology), anti-E-Cadherin antibody (20874-1-AP, Proteintech), anti-Vimentin antibody (#5741, Cell Signaling Technology), anti-PFKFB3 antibody (ab181861, Abcam), anti-LDHA antibody (#3582, Cell Signaling Technology), anti-GLS antibody (ab156876, Abcam), anti-GLUD1 antibody (ab168352, Abcam), anti-PDL1 antibody (#13684S, Cell Signaling Technology), anti-α-actinin antibody (11313-2-AP, Proteintech) and anti-β-actin antibody (#4970, Cell Signaling Technology).

After PET imaging, fluorescence microscopy studies were then used to evaluate PFKFB3 expression in all tumors included in this study. All regents used in this section were purchased from Sigma. Anti-PFKFB3 antibody and goat anti-rabbit IgG H & L (FITC) were purchased from Abcam. A standard immunofluorescence procedure was performed to handle the tumor sections that were removed from the subjects after small PET imaging.