Fig. 5: miR-674-5p targets XBP-1 during LPS stimulation.

a Upper: putative miR-674-5p complementary sequence in the 3′ untranslated region (3′ UTR) of murine XBP-1 mRNA. Lower: conserved miR-674 target sequence in the XBP-1 3′ UTR. b CCC-HIE-2 cells were transfected with a sequence-scrambled oligonucleotide control or miR-674-5p and western blotting was carried out from whole cell lysates collected at various time points. c Luciferase reporter assay was conducted using constructs with the XBP-1 3′ UTR or an antisense control sequence. CCC-HIE-2 cells were cotransfected with these constructs along with the scrambled miRNA or miR-674 mimic. *P < 0.05 versus control 3′ UTR. Three independent experiments were performed. d IEC-6 cells transfected with a scrambled control or anti-miR-674 oligonucleotide were stimulated with LPS, and whole cell lysates were collected at the indicated time points. Three independent experiments were performed. e Quantitative PCR analysis of miR-674-5p gene expression at 24 h in CT-26 cells treated with LPS. *P < 0.05 versus PBS. f CT-26 cells transfected with a scrambled control or anti-miR-674 oligonucleotide were stimulated with LPS, and whole cell lysates were collected at the indicated time points. Western blotting of these lysates is shown.