NheI restriction enzyme

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	The plasmid was digested by NheI (TaKaRa) and HindIII to confirm the correct construction of pcDNA3.1-GLUT1 (Figure 1).

Protocol tips
The plasmid was digested by NheI (TaKaRa) and HindIII to confirm the correct construction of pcDNA3.1-GLUT1 (Figure 1).

Publication protocol

The process of construction of GLUT1 expression vector was in accordance with the protocol described previously [22]. Total RNA was extracted from A549 cells by using RNeasy Mini Kit (Invitrogen) according to the manufacturer’s instructions. The GLUT1 first-strand complementary DNA (cDNA) was generated by using MaximaTM First Strand cDNA Synthesis Kit (Fermentas) and amplified with the primers designed by TaKaRa (Table 1). PCR products and pcDNA3.1 (+) vector (TaKaRa) were digested by HindIII (TaKaRa) and XbaI (TaKaRa). Specific primers (Table 1) were used to subclone the full length coding sequence of GLUT1 into pcDNA3.1 (+) vector to generate pcDNA3.1-GLUT1 (Figure 1). The plasmid was digested by NheI (TaKaRa) and HindIII to confirm the correct construction of pcDNA3.1-GLUT1 (Figure 1).

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