Fig 1: FGF10 is a target of miR-9 in caurulein-treated AR42J cells. (A) Predicted binding sites of miR-9 and FGF10 generated by TargetScan. Binding sites are presented in red and mutant sites are underlined. Luciferase reporter assay was performed in cells co-transfected with (B) wt-FGF10 or mut-FGF10 and miR-NC, miR-9, (C) anti-miR-NC or anti-miR-9. (D) Ago2 RIP assay was performed and miR-9 and FGF10 levels were measured via RT-qPCR. (E) RNA pull-down assay was performed in AR42J cells and miR-9 expression was detected via RT-qPCR. (F) FGF10 expression was detected in cells transfected with miR-NC, miR-9 mimic, anti-miR-NC or anti-miR-9 via western blotting. Data are presented as the mean ± standard deviation. *P<0.05 as indicated. FGF10, fibroblast growth factor 10; wt, wild type; mut, mutant; miR-NC, mimic negative control; miR-9, miR-9 mimic; anti-miR-NC, inhibitor negative control; anti-miR-9, miR-9 inhibitor; Ago2, protein argonaute-2; RIP, RNA immunoprecipitation; RT-qPCR, reverse transcription quantitative PCR; IgG, immunoglobulin G; bio-NC, negative control; bio-FGF10-wt, biotinylated FGF10; bio-FGF10-mut, biotinylated mutant FGF10; input, cell lystates as the positive control.
Fig 2: FGF10 promotes caerulein-induced inflammatory response and apoptosis in AR42J cells. (A) FGF10 expression was measured following treatment at different time points via western blotting. Cells were transfected with pc-NC, pc-FGF10, si-NC or si-FGF10 and (B) FGF10 expression, (C) inflammatory cytokine levels, (D) apoptotic rate, (E) cell apoptosis and (F) apoptotic-associated protein expression were detected via western blotting, ELISA and flow cytometry. Data are presented as the mean ± standard deviation. *P<0.05 as indicated. FGF10, fibroblast growth factor 10; pc-NC, pcDNA empty vector; pc-FGF10, pcDNA-based FGF10 overexpression vector; si-NC, siRNA negative control; si-FGF10, small interfering RNA against FGF10; IL, interleukin; TNF, tumor necrosis factor; Bcl-2, B-cell lymphoma 2; Bax, Bcl-2 associated X; cl-caspase, cleaved caspase.
Fig 3: FGF10 reverses the miR-9-mediated inflammatory response and apoptosis in caerulein-treated AR42J cells. (A) FGF10 expression, inflammatory cytokine (B) IL-1β, (C) IL-6 and (D) TNF-α levels, (E) apoptotic rate and (F) apoptotic-associated protein expressions were detected in cells transfected with miR-9 and pc-NC or pc-FGF10, anti-miR-9 and si-NC or si-FGF10 by ELISA, flow cytometry and western blotting. Data are presented as the mean ± standard deviation. *P<0.05 as indicated. FGF10, fibroblast growth factor 10; miR, microRNA; miR-9, miR-9 mimic; pc-NC, pcDNA empty vector; pc-FGF10, pcDNA-based FGF10 overexpression vector; anti-miR, miR-9 inhibitor; si-NC, siRNA negative control; si-FGF10, siRNA against FGF10; Mock, non-transfected group; IL, interleukin; TNF, tumor necrosis factor; Bcl-2, B-cell lymphoma 2; Bax, Bcl-2 associated X; cl-caspase, cleaved caspase.
Fig 4: miR-9 inhibits the NF-κB pathway through FGF10 in caerulein-treated AR42J cells. (A) IKBα, NFKB1 (p50) and p-p65 expression was measured in AR42J cells transfected with miR-NC, miR-9 mimic, miR-9 mimic and pc-NC or pc-FGF10 after treatment of caerulein by western blotting. (B) IKBα, NF-κB subunit 1 (subunit p50) and phosphorylated p65 expression was measured in AR42J cells transfected with anti-miR-NC, anti-miR-9, anti-miR-9 + si-NC or si-FGF10 following caerulein treatment by western blotting. Data are presented as the mean ± standard deviation. *P<0.05 as indicated. IKBα, NF-kappa-B inhibitor alpha; NF-κB, nuclear factor kappa-light-chain-enhancer of activated B cells; FGF10, fibroblast growth factor 10; p-p65, p-p65; total-p65, p65 in nucleus; anti-miR-NC, inhibitor negative control; pc-NC, pcDNA empty vector; pc-FGF10; pcDNA-based FGF10 overexpression vector; mock, non-transfected group.
Supplier Page from Abcam for Anti-FGF10 antibody