Journal: Breast Cancer : Targets and Therapy

Article Title: Therapeutic Role of Annona muricata Fruit and Bee Venom Against MNU-Induced Breast Cancer in Pregnant Rats and its Complications on the Ovaries

doi: 10.2147/BCTT.S306971

Figure Lengend Snippet: Levels of serum MMP1, NFκB, TNFα and caspase 3 in mother rats and their 21-day offspring among groups. A : The level of serum MMP-1 appears significantly lower in Graviola and/or BV treated mother rats, however this level appears significantly higher in Graviola supplemented offspring if compared with maternally MNU-induced offspring. B and C : A significant decrease in the levels of serum NF-κB and TNF-α in Graviola and/or BV supplemented mother rats and their offspring if compared with MNU-treated group. D : Significant increase in the levels of serum caspase 3 in in Graviola or BV supplemented mother rats and their offspring, however in Graviola and BV supplemented mother rats the level of caspase 3 appears with non-significant change but their offspring display significant increase compared with MNU-treated group.

Article Snippet: NFκB levels in the serum samples were measured using a commercial rat NFκB ELISA kit (E-EL-R0673, Elabscience, China).

Techniques:

Journal: Life

Article Title: Persicaria minor (Huds.) Opiz Prevents In Vitro Atherogenesis by Attenuating Tumor Necrosis Factor-α-Induced Monocyte Adhesion to Human Umbilical Vein Endothelial Cells

doi: 10.3390/life12101462

Figure Lengend Snippet: Nuclear factor kappaB (NF-κB) p65 levels in the nuclear extract of human umbilical vein endothelial cells (HUVEC). Values are expressed as mean ± SEM, n = 7 (* p < 0.05 vs. control. # p < 0.05; ### p < 0.001 vs. TNF-α). PM, standardized aqueous extract of P. minor leaves; Q3G, quercetin-3- O -glucuronide; TNF-α, tumor necrosis factor-α.

Article Snippet: Human NF-κB p65 subunit ELISA kit (Elabscience, Houston, TX, USA) was utilized to determine NF-κB p65 concentration in the nuclear extract of HUVEC.

Techniques: Control

Journal: International Journal of Molecular Sciences

Article Title: In Vitro and In Vivo Validation of Endothelium-Derived Potential Therapeutics for Myocardial Ischemia/Reperfusion Injury Identified by an AI-Enhanced Single-Cell and Virtual-Cell Paradigm

doi: 10.3390/ijms27062743

Figure Lengend Snippet: Andrographolide inhibits the S100A8 downstream IL-17 signaling pathway and reduces the inflammatory response of H/R-induced CMECs. ( A , B ) The expressions of IL-17A, IL-17RA, Act1, and NF-κB proteins were determined by Western blot ( n = 3). ( C – E ) The levels of IL-1β, IL-6, and TNF-α were determined through ELISA. ( F – H ) The expressions of IL-17A, IL-17RA and Act1 proteins were determined by Western blot in co-transfection with the overexpression S100A8 plasmid ( n = 3). Data are presented as mean ± SD ( n = 6). “Con or C” stands for the control group, “H/R” or “H” indicates the H/R group, and “H + AG or H + A” means the H/R group treated with AG. “H + A + S” means the H/R group treated with AG and overexpression of S100A8. ** p < 0.01 compared to the Con group; # p < 0.05, ## p < 0.01 compared to the H/R or H group, @ p < 0.05, @@ p < 0.01 compared to the H + A + S group.

Article Snippet: The membranes were blocked for 1 h at room temperature with 5% skimmed milk before being incubated overnight at 4 °C with the following primary antibodies: S100A8 (A15315, Abclonal, Wuhan, China), IL-17A (A12454, Abclonal, Wuhan, China), IL-17RA (A10052, Abclonal, Wuhan, China), Act1 (A6776, Abclonal, Wuhan, China), p-NF-κB p65 (1:500, AP0124, Abclonal, Wuhan, China), NF-κB p65 (1:500, A00284-1, Boster, Wuhan, China), β-actin (1:2000, TA-09, Zhongshan Golden Bridge, Beijing, China), and GAPDH (1:2000, TA-08, Zhongshan Golden Bridge, Beijing, China).

Techniques: Western Blot, Enzyme-linked Immunosorbent Assay, Cotransfection, Over Expression, Plasmid Preparation, Control

Journal: World Journal of Stem Cells

Article Title: CR6-interacting factor-1 contributes to osteoclastogenesis by inducing receptor activator of nuclear factor κB ligand after radiation

doi: 10.4252/wjsc.v12.i3.222

Figure Lengend Snippet: Radiation induces bone loss and increased CR6-interacting factor-1 expression in mice. A: Micro-computed tomography images of the distal metaphysis of the femur. Mice (n = 6/group ) were exposed to Co-60 gamma rays, and received 5 Gy of whole-body sublethal irradiation at a rate of 0.69 Gy/min; B-H: Micro-computed tomography analysis of the trabecular bone volume/total volume (B), connectivity density (C), trabecular number (D), bone mineral density (E), trabecular thickness (F), trabecular spacing (G), and structure model index (H); I: Hematoxylin-eosin staining of femoral sections from irradiated mice and controls. Femoral sections from irradiated mice showed significantly decreased trabecular bone compared to controls; J: Hematoxylin-eosin staining of femoral sections from irradiated mice and controls. Femoral sections from irradiated mice showed that adipocytes increased significantly in irradiated mice; K: Tartrate-resistant acid phosphatase staining of femoral sections from irradiated mice and controls; L: Real-time quantitative polymerase chain reaction (RT-qPCR) analysis of receptor activator of nuclear factor κB ligand and osteoprotegerin mRNA expression in flushed whole bone marrow; M: Receptor activator of nuclear factor κB ligand/osteoprotegerin ratio based on RT-qPCR results; N: RT-qPCR analysis of CR6-interacting factor-1 mRNA expression in flushed whole bone marrow; O: Western blot analysis of CR6-interacting factor-1 expression in flushed whole bone marrow. aP < 0.05, bP < 0.01, and the bars represent the mean ± SD. OPG: Osteoprotegerin; RANKL: Receptor activator of nuclear factor κB ligand; Crif1: CR6-interacting factor-1; BM: Bone marrow; SMI: Structure model index; BV/TV: Bone volume/total volume; Conn.D: Connectivity density; Tb.N: Trabecular number; vBMD: Bone mineral density; Tb.Sp: Trabecular spacing; Tb.Th: Trabecular thickness.

Article Snippet: ELISA The concentrations of RANKL and OPG were measured using the Mouse RANKL ELISA Kit (E-EL-M0644c, elabscience), Human Soluble Receptor Activator of Nuclear Factor-kB Ligand ELISA Kit (E-EL-H5558c, elabscience), Mouse OPG ELISA Kit (E-EL-M0081c, elabscience), and Human OPG ELISA Kit (E-EL-H1341c, elabscience) according to the manufacturer’s instructions.

Techniques: Expressing, Micro-CT, Irradiation, Staining, Real-time Polymerase Chain Reaction, Quantitative RT-PCR, Western Blot

Journal: World Journal of Stem Cells

Article Title: CR6-interacting factor-1 contributes to osteoclastogenesis by inducing receptor activator of nuclear factor κB ligand after radiation

doi: 10.4252/wjsc.v12.i3.222

Figure Lengend Snippet: Overexpression of CR6-interacting factor-1 in bone marrow mesenchymal stem/stromal cells increases receptor activator of nuclear factor κB ligand secretion and osteoclastogenesis. A: Western blot analysis of CR6-interacting factor-1 (Crif1) expression in mouse bone marrow mesenchymal stem/stromal cells (BM-MSCs). Mouse BM-MSCs were transfected with a Crif1 lentiviral overexpression vector; B: Real-time quantitative polymerase chain reaction analysis of receptor activator of nuclear factor κB ligand (RANKL) and osteoprotegerin (OPG) mRNA expression in BM-MSCs and Crif1-overexpressing BM-MSCs. BM-MSCs and Crif1-overexpressing BM-MSCs were cocultured with RAW264.7, respectively; C: RANKL/OPG ratio based on real-time quantitative polymerase chain reaction results; D: Enzyme linked immunosorbent assay analysis of RANKL protein levels in coculture supernatant medium; E: Enzyme linked immunosorbent assay analysis of OPG protein levels in coculture supernatant medium; F: RANKL/OPG ratio in coculture supernatant medium; G: Tartrate-resistant acid phosphatase staining of RAW264.7 cells after 7 d of coculture; H: Average number of tartrate-resistant acid phosphatase-positive cells/well (arrow) from RAW264.7 cells in coculture. aP < 0.05, bP < 0.01, and the bars represent the mean ± SD. OPG: Osteoprotegerin; RANKL: Receptor activator of nuclear factor κB ligand; Crif1: CR6-interacting factor-1; BM-MSCs: Bone marrow mesenchymal stem/stromal cells; BM-MSCs-OV: Crif1-overexpressing BM-MSCs.

Article Snippet: ELISA The concentrations of RANKL and OPG were measured using the Mouse RANKL ELISA Kit (E-EL-M0644c, elabscience), Human Soluble Receptor Activator of Nuclear Factor-kB Ligand ELISA Kit (E-EL-H5558c, elabscience), Mouse OPG ELISA Kit (E-EL-M0081c, elabscience), and Human OPG ELISA Kit (E-EL-H1341c, elabscience) according to the manufacturer’s instructions.

Techniques: Over Expression, Western Blot, Expressing, Transfection, Plasmid Preparation, Real-time Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay, Staining

Journal: World Journal of Stem Cells

Article Title: CR6-interacting factor-1 contributes to osteoclastogenesis by inducing receptor activator of nuclear factor κB ligand after radiation

doi: 10.4252/wjsc.v12.i3.222

Figure Lengend Snippet: CR6-interacting factor-1 mediates adipogenesis and receptor activator of nuclear factor κB ligand secretion in adipocytes. A: Oil red O staining analysis of mouse bone marrow mesenchymal stem/stromal cells (BM-MSCs) after 21 d of adipogenic differentiation. Crif1 was knocked out in mouse BM-MSCs (BM-MSCs-KO), and knockout cells and controls were irradiated with 9 Gy of Co-60, and then treated with mouse mesenchymal stem cell adipogenic differentiation medium (Ad) to induce adipogenesis; B: The dye from oil red O staining was extracted using isopropanol, and the optical density at 510 nm was measured using Benchmark Plus; C: Western blot analysis of adipogenesis-related markers and transcription factors PPARγ and AP2 in mouse BM-MSCs after 21 d of adipogenic differentiation; D: Real-time quantitative polymerase chain reaction analysis of receptor activator of nuclear factor κB ligand (RANKL) and osteoprotegerin (OPG) mRNA expression in BM-MSCs and BM-MSCs-KO; E: RANKL/OPG ratio based on real-time quantitative polymerase chain reaction results; F: Enzyme linked immunosorbent assay analysis of RANKL protein levels in supernatant Ad; G: Enzyme linked immunosorbent assay analysis of OPG protein levels in supernatant Ad; H: RANKL/OPG ratio in supernatant Ad. aP < 0.05 vs control (BM-MSCs), bP < 0.01 vs control (BM-MSCs); dP < 0.01 between 9 Gy-BM-MSCs and 9 Gy-BM-MSCs-KO, and the bars represent the mean ± SD. BM-MSCs: Bone marrow mesenchymal stem/stromal cells; BM-MSCs-KO: Crif1 knockout mouse BM-MSCs; OPG: Osteoprotegerin; RANKL: Receptor activator of nuclear factor κB ligand.

Article Snippet: ELISA The concentrations of RANKL and OPG were measured using the Mouse RANKL ELISA Kit (E-EL-M0644c, elabscience), Human Soluble Receptor Activator of Nuclear Factor-kB Ligand ELISA Kit (E-EL-H5558c, elabscience), Mouse OPG ELISA Kit (E-EL-M0081c, elabscience), and Human OPG ELISA Kit (E-EL-H1341c, elabscience) according to the manufacturer’s instructions.

Techniques: Staining, Knock-Out, Irradiation, Western Blot, Real-time Polymerase Chain Reaction, Expressing, Enzyme-linked Immunosorbent Assay, Control

Journal: World Journal of Stem Cells

Article Title: CR6-interacting factor-1 contributes to osteoclastogenesis by inducing receptor activator of nuclear factor κB ligand after radiation

doi: 10.4252/wjsc.v12.i3.222

Figure Lengend Snippet: CR6-interacting factor-1 is involved in the regulation of receptor activator of nuclear factor κB ligand expression after radiation. A: Western blot analysis of CR6-interacting factor-1 (Crif1) and receptor activator of nuclear factor κB expression in RAW264.7 cells. Crif1 was knocked out in RAW264.7 cells (RAW264.7-KO); B: Tartrate-resistant acid phosphatase (TRAP) staining of RAW264.7-KO and controls after 7 d of coculture with mouse bone marrow mesenchymal stem/stromal cells (BM-MSCs); C: Average number of TRAP-positive cells/well (arrow) from RAW264.7-KO and controls after 7 d of coculture with mouse BM-MSCs; D: Western blot analysis of Crif1 expression in BM-MSCs. Crif1 was knocked out in mouse BM-MSCs (BM-MSCs-KO), and BM-MSCs-KO and controls were irradiated with Co-60 at a single dose of 9 Gy; E: Real-time quantitative polymerase chain reaction analysis of receptor activator of nuclear factor κB ligand (RANKL) and osteoprotegerin (OPG) mRNA expression in BM-MSCs and BM-MSCs-KO. BM-MSCs and BM-MSCs-KO were cocultured with RAW264.7; F: RANKL/OPG ratio based on real-time quantitative polymerase chain reaction results; G: Enzyme linked immunosorbent assay analysis of RANKL protein levels in coculture supernatant medium; H: Enzyme linked immunosorbent assay analysis of OPG protein levels in coculture supernatant medium; I: RANKL/OPG ratio in coculture supernatant medium; J: TRAP staining of RAW264.7 after 7 d of coculture; K: Average number of TRAP-positive cells/well (arrow) from RAW264.7 in coculture. aP < 0.05 vs control (BM-MSCs), bP < 0.01 vs control (BM-MSCs); dP < 0.01 between 9 Gy-BM-MSCs and 9 Gy-BM-MSCs-KO, and the bars represent the mean ± standard deviation. BM-MSCs: Bone marrow mesenchymal stem/stromal cells; BM-MSCs-KO: Crif1 was knocked out from mouse BM-MSCs; RAW264.7-KO: Crif1 was knocked out from RAW264.7 cells; OPG: Osteoprotegerin; RANKL: Receptor activator of nuclear factor κB ligand.

Article Snippet: ELISA The concentrations of RANKL and OPG were measured using the Mouse RANKL ELISA Kit (E-EL-M0644c, elabscience), Human Soluble Receptor Activator of Nuclear Factor-kB Ligand ELISA Kit (E-EL-H5558c, elabscience), Mouse OPG ELISA Kit (E-EL-M0081c, elabscience), and Human OPG ELISA Kit (E-EL-H1341c, elabscience) according to the manufacturer’s instructions.

Techniques: Expressing, Western Blot, Staining, Irradiation, Real-time Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay, Control, Standard Deviation

Journal: World Journal of Stem Cells

Article Title: CR6-interacting factor-1 contributes to osteoclastogenesis by inducing receptor activator of nuclear factor κB ligand after radiation

doi: 10.4252/wjsc.v12.i3.222

Figure Lengend Snippet: CR6-interacting factor-1 promotes receptor activator of nuclear factor κB ligand secretion by modulating the cAMP/PKA signaling pathway. A: Real-time quantitative polymerase chain reaction (RT-qPCR) analysis of receptor activator of nuclear factor κB ligand (RANKL) and osteoprotegerin (OPG) mRNA expression in bone marrow mesenchymal stem/stromal cells (BM-MSCs) and Crif1 knockout BM-MSCs treated with 25 µmol/L forskolin in the coculture with RAW264.7; B: RANKL/OPG ratio based on RT-qPCR results; C: Enzyme linked immunosorbent assay (ELISA) analysis of RANKL levels in coculture supernatant medium treated with 25 µmol/L forskolin; D: ELISA analysis of OPG levels in coculture supernatant medium treated with 25 µmol/L forskolin; E: RANKL/OPG ratio in coculture supernatant medium treated with 25 µmol/L forskolin; F: RT-qPCR analysis of RANKL and OPG mRNA expression in BM-MSCs and BM-MSCs-OV treated with 20 µmol/L H-89 in the coculture with RAW264.7; G: RANKL/OPG ratio based on RT-qPCR results; H: ELISA analysis of RANKL levels in coculture supernatant medium treated with 20 µmol/L H-89; I: ELISA analysis of OPG levels in coculture supernatant medium treated with 20 µmol/L H-89; J: RANKL/OPG ratio in coculture supernatant medium treated with 20 µmol/L H-89; K: Tartrate-resistant acid phosphatase (TRAP) staining of RAW264.7 cells in coculture treated with 25 µmol/L forskolin; L: Average number of TRAP-positive cells/well (arrow) from RAW264.7 cells in coculture treated with 25 µmol/L forskolin; M: Western blot analysis of phospho-cyclic adenosine monophosphate response element-binding protein phosphorylation levels in BM-MSCs in coculture treated with 25 µmol/L forskolin; N: TRAP staining of RAW264.7 in coculture treated with 20 µmol/L H-89; O: Average number of TRAP-positive cells/well (arrow) from RAW264.7 in coculture treated with 20 µmol/L H-89; P: Western blot analysis of phospho-cyclic adenosine monophosphate response element-binding protein phosphorylation levels in BM-MSCs in coculture treated with 20 µmol/L H-89. bP < 0.01 vs control (BM-MSCs); dP < 0.01 between between BM-MSCs treated with 25 µmol/L forskolin and Crif1 knockout BM-MSCs treated with 25 µmol/L forskolin; fP < 0.01 between Crif1-overexpressing BM-MSCs and Crif1-overexpressing BM-MSCs treated with 20 µmol/L H-89, and the bars represent the mean ± SD. BM-MSCs: Bone marrow mesenchymal stem/stromal cells; BM-MSCs-KO: Crif1 knockout BM-MSCs; OPG: Osteoprotegerin; RANKL: Receptor activator of nuclear factor κB ligand; CREB: Cyclic adenosine monophosphate response element-binding protein.

Article Snippet: ELISA The concentrations of RANKL and OPG were measured using the Mouse RANKL ELISA Kit (E-EL-M0644c, elabscience), Human Soluble Receptor Activator of Nuclear Factor-kB Ligand ELISA Kit (E-EL-H5558c, elabscience), Mouse OPG ELISA Kit (E-EL-M0081c, elabscience), and Human OPG ELISA Kit (E-EL-H1341c, elabscience) according to the manufacturer’s instructions.

Techniques: Real-time Polymerase Chain Reaction, Quantitative RT-PCR, Expressing, Knock-Out, Enzyme-linked Immunosorbent Assay, Staining, Western Blot, Binding Assay, Phospho-proteomics, Control

Journal: World Journal of Stem Cells

Article Title: CR6-interacting factor-1 contributes to osteoclastogenesis by inducing receptor activator of nuclear factor κB ligand after radiation

doi: 10.4252/wjsc.v12.i3.222

Figure Lengend Snippet: CR6-interacting factor-1 inhibitors effectively suppress receptor activator of nuclear factor κB ligand secretion and adipogenesis. A: Enzyme linked immunosorbent assay (ELISA) analysis of receptor activator of nuclear factor κB ligand protein levels in the supernatant medium. Human bone marrow mesenchymal stem/stromal cells (H-BM-MSCs) were pretreated with five different compounds (25 µmol/L) followed by treatment with forskolin (25 µmol/L), and supernatant medium was collected for ELISA after 3 d; B: ELISA analysis of osteoprotegerin protein levels in supernatant medium; C: Receptor activator of nuclear factor κB ligand/osteoprotegerin ratio in supernatant medium; D: Oil red O staining analysis of H-BM-MSCs after 21 d of adipogenic differentiation. H-BM-MSCs were pretreated with five different compounds (25 µmol/L) followed by adipogenic induction; E: The dye from oil red O staining was extracted using isopropanol, and the optical density at 510 nm was measured using Benchmark Plus; F: Western blot analysis of cyclic adenosine monophosphate response element-binding protein phosphorylation levels. H-BM-MSCs were pretreated with five different compounds (25 µmol/L) followed by treatment with forskolin (25 µmol/L) and total protein lysates were extracted for cyclic adenosine monophosphate response element-binding protein phosphorylation detection after 1 h. aP < 0.05, bP < 0.01, and the bars represent the mean ± SD. OPG: Osteoprotegerin; RANKL: Receptor activator of nuclear factor κB ligand; CREB: Cyclic adenosine monophosphate response element-binding protein; H-BM-MSCs: Human bone marrow mesenchymal stem/stromal cells.

Article Snippet: ELISA The concentrations of RANKL and OPG were measured using the Mouse RANKL ELISA Kit (E-EL-M0644c, elabscience), Human Soluble Receptor Activator of Nuclear Factor-kB Ligand ELISA Kit (E-EL-H5558c, elabscience), Mouse OPG ELISA Kit (E-EL-M0081c, elabscience), and Human OPG ELISA Kit (E-EL-H1341c, elabscience) according to the manufacturer’s instructions.

Techniques: Enzyme-linked Immunosorbent Assay, Staining, Western Blot, Binding Assay, Phospho-proteomics