Journal: Oncotarget

Article Title: IL-1β-stimulated β-catenin up-regulation promotes angiogenesis in human lung-derived mesenchymal stromal cells through a NF-κB-dependent microRNA-433 induction

doi: 10.18632/oncotarget.10683

Figure Lengend Snippet: A. Levels of miR-433 in hL-MSC after IL-1β treatment, with PBS as control. B. mRNA levels of genes known to be inhibited by IL-1β after IL-1β treatment, with PBS as control. C. mRNA levels of genes in hL-MSC transfected with either miR-negative control (NC) or miR-433. Values were mean ± SD from three independent experiments. ** P < 0.01, * P < 0.05 vs PBS or miR-NC, respectively.

Article Snippet: The mir Vana miRNA Isolation Kit (AM1561, Applied Biosystems) was used to isolate total miRNA, and expression levels of miR-433 were then determined by pri-miRNA assay kit (Hs03303744_pri, Applied Biosystems) and mature miRNA assay kit (478102_mir, Applied Biosystems) according to manufacturer's instructions.

Techniques: Control, Transfection, Negative Control

Journal: Oncotarget

Article Title: IL-1β-stimulated β-catenin up-regulation promotes angiogenesis in human lung-derived mesenchymal stromal cells through a NF-κB-dependent microRNA-433 induction

doi: 10.18632/oncotarget.10683

Figure Lengend Snippet: A. and B. Wound healing (A) and tube formation (B) assays were performed in hL-MSC derived endothelial cells treated with PBS or IL-1β. C. and D. Wound healing (C) and tube formation (D) assays were performed in hL-MSC derived endothelial cells transfected with miR-NC or miR-433. E. and F. hL-MSC derived endothelial cells treated with PBS or IL-1β were also transfected with either miR-NC or anti-miR-433, followed by wound healing (E) and tube formation (F) assays to assess their angiogenic capacity. Values were mean ± SD from three independent experiments. ** P < 0.01, * P < 0.05, ns not significant vs respective control.

Article Snippet: The mir Vana miRNA Isolation Kit (AM1561, Applied Biosystems) was used to isolate total miRNA, and expression levels of miR-433 were then determined by pri-miRNA assay kit (Hs03303744_pri, Applied Biosystems) and mature miRNA assay kit (478102_mir, Applied Biosystems) according to manufacturer's instructions.

Techniques: Derivative Assay, Transfection, Control

Journal: Oncotarget

Article Title: IL-1β-stimulated β-catenin up-regulation promotes angiogenesis in human lung-derived mesenchymal stromal cells through a NF-κB-dependent microRNA-433 induction

doi: 10.18632/oncotarget.10683

Figure Lengend Snippet: A. Sequence of the putative miR-433 targeting site (capitalized) on the 3′-UTR of DKK1 mRNA. B. Wild type (-Wt) or mutated (-Mut) versions of putative targeting sequence from the 3′-UTR of DKK1 mRNA were fused after the downstream of a luciferase reporter (Luc) open reading frame. C. and D. Luciferase activities of Luc-Wt and Luc-Mut constructs were measured in hL-MSC after transfection with either miR-NC or miR-433 (C), or treatment with either PBS or IL-1β (D). E. DKK1 protein levels in hL-MSC after transfection with either miR-NC or miR-433. F. hL-MSC treated with PBS or IL-1β were also transfected with either miR-NC or miR-433 inhibitor (anti-miR-433), followed by Western blot analysis to examine DKK1 protein levels. Values were mean ± SD from three independent experiments. ** P < 0.01, * P < 0.05, ns not significant vs PBS or miR-NC, respectively.

Article Snippet: The mir Vana miRNA Isolation Kit (AM1561, Applied Biosystems) was used to isolate total miRNA, and expression levels of miR-433 were then determined by pri-miRNA assay kit (Hs03303744_pri, Applied Biosystems) and mature miRNA assay kit (478102_mir, Applied Biosystems) according to manufacturer's instructions.

Techniques: Sequencing, Luciferase, Construct, Transfection, Western Blot

Journal: Oncotarget

Article Title: IL-1β-stimulated β-catenin up-regulation promotes angiogenesis in human lung-derived mesenchymal stromal cells through a NF-κB-dependent microRNA-433 induction

doi: 10.18632/oncotarget.10683

Figure Lengend Snippet: A. Levels of miR-433 in the presence of IL-1β were examined in hL-MSC after treatment with either NF-кB inhibitor TPCA-1, p38 inhibitor BIX02188 or JNK inhibitor SP600125, respectively. B. and C. Levels of NF-кB protein (B) and miR-433 (C) in hL-MSC following NF-кB siRNA knock-down were examined by Western blot and RT-PCR, respectively. D. Protein levels of DKK1 in hL-MSC in the presence of IL-1β were examined by Western blot, following treatment with the inhibitors used in (A) or NF-кB siRNA knock-down used in (B). Values were mean ± SD from three independent experiments. ** P < 0.01, ns not significant vs mock or siRNA NC, respectively.

Article Snippet: The mir Vana miRNA Isolation Kit (AM1561, Applied Biosystems) was used to isolate total miRNA, and expression levels of miR-433 were then determined by pri-miRNA assay kit (Hs03303744_pri, Applied Biosystems) and mature miRNA assay kit (478102_mir, Applied Biosystems) according to manufacturer's instructions.

Techniques: Knockdown, Western Blot, Reverse Transcription Polymerase Chain Reaction

Journal: Oncotarget

Article Title: IL-1β-stimulated β-catenin up-regulation promotes angiogenesis in human lung-derived mesenchymal stromal cells through a NF-κB-dependent microRNA-433 induction

doi: 10.18632/oncotarget.10683

Figure Lengend Snippet: A. Promoter region of human miR-433 contains two putative binding sites for NF-κB, which was then clone to the upstream of a luciferase reporter (Luc) open reading frame. B. Binding of NF-κB to the promoter of miR-433 in hL-MSC was examined by ChIP assay using control IgG or NF-κB antibody. C. Luciferase activities of AB-Luc, A-Luc and B-Luc constructs were measured in hL-MSC after treatment with either PBS or NF-κB. Values were mean ± SD from three independent experiments. ** P < 0.01, ns not significant vs control IgG or PBS, respectively.

Article Snippet: The mir Vana miRNA Isolation Kit (AM1561, Applied Biosystems) was used to isolate total miRNA, and expression levels of miR-433 were then determined by pri-miRNA assay kit (Hs03303744_pri, Applied Biosystems) and mature miRNA assay kit (478102_mir, Applied Biosystems) according to manufacturer's instructions.

Techniques: Binding Assay, Luciferase, Control, Construct

Journal: Oncotarget

Article Title: IL-1β-stimulated β-catenin up-regulation promotes angiogenesis in human lung-derived mesenchymal stromal cells through a NF-κB-dependent microRNA-433 induction

doi: 10.18632/oncotarget.10683

Figure Lengend Snippet: A. Levels of β-catenin mRNA in hL-MSC transfected with either miR-NC or miR-433. B. hL-MSC treated with PBS or IL-1β were also transfected with either miR-NC or anti-miR-433, followed by RT-PCR analysis to examine β-catenin mRNA levels. C. Levels of β-catenin mRNA in hL-MSC treated with PBS, IL-1β or IL-1β + NF-кB inhibitor TPCA-1. Values were mean ± SD from three independent experiments. ** P < 0.01, * P < 0.05, ns not significant vs miR-NC or PBS, respectively. D. A schematic diagram illustrating the mechanism of IL-1β-stimulated β-catenin up-regulation, mediated by NF-κb-dependent miRNA-433 induction, to promote angiogenesis in hL-MSC.

Article Snippet: The mir Vana miRNA Isolation Kit (AM1561, Applied Biosystems) was used to isolate total miRNA, and expression levels of miR-433 were then determined by pri-miRNA assay kit (Hs03303744_pri, Applied Biosystems) and mature miRNA assay kit (478102_mir, Applied Biosystems) according to manufacturer's instructions.

Techniques: Transfection, Reverse Transcription Polymerase Chain Reaction