RNA interferences
The siRNAs for ADAM17 and 11β-HSD2 were designed and synthesized by
GenePharma Corporation (Shanghai, China). Control siRNA was scrambled
sequence without any specific target. The sequences for targeting human
negative control are: sense: 5’-UUCUCCGAACGUGUCACGUTT-3’, antisense:
5’-ACGUGACACGUUCGGAGAATT-3’. The sequences for targeting human ADAM17
are: sense: 5’-CACAUGUAGAAACACUACUTT -3’, antisense:
5’-AGUAGUGUUUCUACAUGUIGTT-3’. The sequences for targeting human 11β-HSD2
are: sense: 5’-CCACUUUCCGUAGCUGCAUTT-3’, antisense:
5’-AUGCAGCUACGGAAAGUGGTT-3’; Transfection of siRNA was performed by
using Lipofectamine TM 3000 (Invitrogen, Thermo Fisher
Scientific, Rockford, IL) as described previously
(He et al., 2014). The efficiency of
ADAM17 and 11β-HSD2 were determined by western blotting.