Quantifying Glucose Transporter Expression

After drug treatment, mRNA was obtained from cortical or hippocampal tissue and used to generate cDNA. Quantitative real-time RT–PCR (qRT–PCR) was conducted using SYBR master mix (#4368577, ThermoFisher Scientific), with the program recommended by the manufacturer and as published previously (38 (link)). As a reference, we used the housekeeping gene cyclophilin (Ppib), and the relative Ct values of each gene were calculated using the delta Ct, in comparison with the control gene. Duplicated control reactions for every sample without reverse transcription were included to ensure that PCR products were not due to the amplification of contaminated genomic DNA. We used the following sets of primers: cyclophilin F (5’-TGGAGATGAATCTGTAGGAGGAG-3’) and R (5’- TACCACATCCATGCCCTCTAGAA-3), Glut1 (Slc2a1) F (5’-ATGGATCCCAGCAGCA AGAAG-3’) and R (5’-AGAGACCAAAGCGTGGTGAG-3’), Glut3 (Slc2a3) F (5’-GGATCCCTTGTCCTTCTGCTT-3’) and R (5’-ACCAGTTCCCAATGCACACA-3’), Glut4 (Slc2a4) F (5’-CGGCTCTGACGATGGGGAA-3’) and R (5’-TTGTGGGATGGAA TCCGGTCCCGATA-3’). All primers were purchased from IDT Integrated DNA Technologies.