Figure 5.

miR-1 and miR-122 reppress G6PD expression by direct targeting. (a) G6PD was validated as a miR-1 and miR-122 target using luciferase reporter assay. H293-T cells were transfected with control RNA (NC), miR-122, miR-1, or combination of both, along with psi-CHECK2 vector harboring human G6PD 3′-UTR and Firefly luciferase (an internal control). After 48 hours, luciferase activities were measured in cell extracts per manufacture’s protocol (Promega). (b,c) G6PD immunoblot and RT-qPCR analysis shows depletion of G6PD protein level (b) and mRNA level (c) in HCC cells transfected with miRNA mimics (50 nM) for 48hrs. (* indicates p-value < 0.05, p-value was calculated using Student T-test). Cropped immunoblot images in (b) were obtained from the same gel. Full immunoblot images are found in supplemental material.