Additional file 4: of Isoliquiritigenin suppresses human melanoma growth by targeting miR-301b/LRIG1 signaling

Figure S3. (A)RT-qPCR analysis of the mRNA level alteration of 7 common target genes of miR301b in A375 and A2058 cells after being transfected with miR301b mimic/NC or treated with miR301b inhibitor/NC. *P < 0.05 vs NC. (B)Design of luciferase reporters with the WT Akt3 3’UTR (Akt3–3’UTR WT) or the site-directed mutant Akt3 3’UTR (Akt3–3’UTR MUT). (C)RT-qPCR analysis of miR301b level in A375 and A2058 cells after being transfected with miR301b mimic/NC or treated with miR301b inhibitor/NC. **P < 0.01 vs NC. (D)RT-qPCR analysis of the mRNA level of LRIG1 in A375 and A2058 cells after being transfected with miR301b mimic/NC or treated with miR301b inhibitor/NC. **P < 0.01 vs NC.*P < 0.05 vs NC. (E, F)Western blot analysis of the protein level of apoptosis associated proteins(LRIG1, c-PARP, Bax, cleaved-caspase-3) in ISL treated A375 and A2058 cells which were transfected with si-LRIG1 or control(NC). *P < 0.05, **P < 0.01 vs PBS Treated in si-NC groups. #P < 0.05, ##P < 0.01 vs PBS Treated in si-LRIG1 groups. (G)Flow cytometry analysis of cell apoptosis in ISL treated A375 and A2058 cells which were transfected with si-LRIG1 or si-NC.*P < 0.05, **P < 0.01 vs PBS + si-NC. (H)Quantification of TUNEL positive cells in ISL treated A375 and A2058 cells which were transfected with si-LRIG1 or si-NC.*P < 0.05, **P < 0.01 vs PBS + si-NC. (TIF 25520 kb