Journal: International Journal of Molecular Medicine

Article Title: MicroRNA-155 inhibits the osteogenic differentiation of mesenchymal stem cells induced by BMP9 via downregulation of BMP signaling pathway

doi: 10.3892/ijmm.2018.3526

Figure Lengend Snippet: Primers used in the study.

Article Snippet: Specific protein bands were detected with mouse anti-BMPR2 polyclonal antibody (1:200 dilution, cat. no. BM2821; BOSTER), rabbit anti-p-Smad1/5/8 monoclonal antibody (1:1,000 dilution, cat. no. sc-12353), rabbit anti-Smad1/5/8 monoclonal antibody (1:1,000 dilution, cat. no. sc-6031-R, lot no. D1911), rabbit anti-Runx2 monoclonal antibody (1:1,000 dilution, cat. no. sc-10758, lot no. A0810) (all from Santa Cruz Biotechnology, Inc., Santa Cruz, CA, USA), rabbit anti-osteocalcin (OCN) polyclonal antibody (1:200 dilution, cat. no. bs-4917R; Beijing Biosynthesis Biotechnology Co., Ltd.), rabbit anti-osteopontin (OPN) polyclonal antibody (1:500 dilution, cat. no. wl00691; Wanleibio, Shenyang, China), or anti-β-actin mouse monoclonal antibody (1:1,000 dilution; Zhongshan Golden Bridge Biotechnology Co., Ltd., Beijing, China) overnight at 4°C, and then washed with TBST three times, each time for 15 min, the membranes were incubated with goat anti-rabbit or goat anti-mouse horseradish peroxidase conjugated secondary antibody (1:5,000 dilution; Zhongshan Golden Bridge Biotechnology Co., Ltd.), for 1 h at 37°C.

Techniques: Sequencing

Journal: International Journal of Molecular Medicine

Article Title: MicroRNA-155 inhibits the osteogenic differentiation of mesenchymal stem cells induced by BMP9 via downregulation of BMP signaling pathway

doi: 10.3892/ijmm.2018.3526

Figure Lengend Snippet: The sequences contained binding sites with miR-155.

Article Snippet: Specific protein bands were detected with mouse anti-BMPR2 polyclonal antibody (1:200 dilution, cat. no. BM2821; BOSTER), rabbit anti-p-Smad1/5/8 monoclonal antibody (1:1,000 dilution, cat. no. sc-12353), rabbit anti-Smad1/5/8 monoclonal antibody (1:1,000 dilution, cat. no. sc-6031-R, lot no. D1911), rabbit anti-Runx2 monoclonal antibody (1:1,000 dilution, cat. no. sc-10758, lot no. A0810) (all from Santa Cruz Biotechnology, Inc., Santa Cruz, CA, USA), rabbit anti-osteocalcin (OCN) polyclonal antibody (1:200 dilution, cat. no. bs-4917R; Beijing Biosynthesis Biotechnology Co., Ltd.), rabbit anti-osteopontin (OPN) polyclonal antibody (1:500 dilution, cat. no. wl00691; Wanleibio, Shenyang, China), or anti-β-actin mouse monoclonal antibody (1:1,000 dilution; Zhongshan Golden Bridge Biotechnology Co., Ltd., Beijing, China) overnight at 4°C, and then washed with TBST three times, each time for 15 min, the membranes were incubated with goat anti-rabbit or goat anti-mouse horseradish peroxidase conjugated secondary antibody (1:5,000 dilution; Zhongshan Golden Bridge Biotechnology Co., Ltd.), for 1 h at 37°C.

Techniques: Binding Assay, Sequencing

Journal: International Journal of Molecular Medicine

Article Title: MicroRNA-155 inhibits the osteogenic differentiation of mesenchymal stem cells induced by BMP9 via downregulation of BMP signaling pathway

doi: 10.3892/ijmm.2018.3526

Figure Lengend Snippet: The expression level of miR-155 is increased at first and then decreased in the process of bone morphogenetic protein 9 (BMP9)-induced early osteogenesis. (A) The expression of miR-155 was detected by RT-qPCR. * p<0.05 and *** p<0.001 vs. the value at day 0. (B) BMP9 was transfected into C2C12 cells and mouse embryonic fibroblasts (MEF) cells efficiently. Magnification, ×100. White, white light used as the excitation light; green, blue light used as the excitation light. Multiple of infection (MOI): 5. (C) The expression of runt-related transcription factor 2 (Runx2) and alkaline phosphatase (ALP) in C2C12 cells and MEF cells in differentiation days induced by BMP9 were detected by RT-qPCR. ** p<0.01 and *** p<0.001 vs. the value at day 0. Day: osteogenic differentiation day.

Article Snippet: Specific protein bands were detected with mouse anti-BMPR2 polyclonal antibody (1:200 dilution, cat. no. BM2821; BOSTER), rabbit anti-p-Smad1/5/8 monoclonal antibody (1:1,000 dilution, cat. no. sc-12353), rabbit anti-Smad1/5/8 monoclonal antibody (1:1,000 dilution, cat. no. sc-6031-R, lot no. D1911), rabbit anti-Runx2 monoclonal antibody (1:1,000 dilution, cat. no. sc-10758, lot no. A0810) (all from Santa Cruz Biotechnology, Inc., Santa Cruz, CA, USA), rabbit anti-osteocalcin (OCN) polyclonal antibody (1:200 dilution, cat. no. bs-4917R; Beijing Biosynthesis Biotechnology Co., Ltd.), rabbit anti-osteopontin (OPN) polyclonal antibody (1:500 dilution, cat. no. wl00691; Wanleibio, Shenyang, China), or anti-β-actin mouse monoclonal antibody (1:1,000 dilution; Zhongshan Golden Bridge Biotechnology Co., Ltd., Beijing, China) overnight at 4°C, and then washed with TBST three times, each time for 15 min, the membranes were incubated with goat anti-rabbit or goat anti-mouse horseradish peroxidase conjugated secondary antibody (1:5,000 dilution; Zhongshan Golden Bridge Biotechnology Co., Ltd.), for 1 h at 37°C.

Techniques: Expressing, Quantitative RT-PCR, Transfection, Infection

Journal: International Journal of Molecular Medicine

Article Title: MicroRNA-155 inhibits the osteogenic differentiation of mesenchymal stem cells induced by BMP9 via downregulation of BMP signaling pathway

doi: 10.3892/ijmm.2018.3526

Figure Lengend Snippet: miR-155 downregulates the mRNA expression levels of osteogenesis genes runt-related transcription factor 2 (Runx2), osterix (OSX), alkaline phosphatase (ALP) and osteocalcin (OCN). C2C12 cells and mouse embryonic fibroblasts (MEF) were transfected with NC, miR-155 or anti-miR-155, respectively, then induced osteogenesis by bone morphogenetic protein 9 (BMP9). (A) The expression of miR-155 by RT-qPCR in the third day of differentiation. (B) Runx2 and OSX were tested on the 3rd day of differentiation, ALP and OCN were tested on the 7th day of differentiation in C2C12 cells. (C) Runx2 and OSX were tested on the 3rd day of differentiation, ALP and OCN were tested on the 7th day of differentiation in MEF cells. * p<0.05, ** p<0.01 and *** p<0.001.

Article Snippet: Specific protein bands were detected with mouse anti-BMPR2 polyclonal antibody (1:200 dilution, cat. no. BM2821; BOSTER), rabbit anti-p-Smad1/5/8 monoclonal antibody (1:1,000 dilution, cat. no. sc-12353), rabbit anti-Smad1/5/8 monoclonal antibody (1:1,000 dilution, cat. no. sc-6031-R, lot no. D1911), rabbit anti-Runx2 monoclonal antibody (1:1,000 dilution, cat. no. sc-10758, lot no. A0810) (all from Santa Cruz Biotechnology, Inc., Santa Cruz, CA, USA), rabbit anti-osteocalcin (OCN) polyclonal antibody (1:200 dilution, cat. no. bs-4917R; Beijing Biosynthesis Biotechnology Co., Ltd.), rabbit anti-osteopontin (OPN) polyclonal antibody (1:500 dilution, cat. no. wl00691; Wanleibio, Shenyang, China), or anti-β-actin mouse monoclonal antibody (1:1,000 dilution; Zhongshan Golden Bridge Biotechnology Co., Ltd., Beijing, China) overnight at 4°C, and then washed with TBST three times, each time for 15 min, the membranes were incubated with goat anti-rabbit or goat anti-mouse horseradish peroxidase conjugated secondary antibody (1:5,000 dilution; Zhongshan Golden Bridge Biotechnology Co., Ltd.), for 1 h at 37°C.

Techniques: Expressing, Transfection, Quantitative RT-PCR

Journal: International Journal of Molecular Medicine

Article Title: MicroRNA-155 inhibits the osteogenic differentiation of mesenchymal stem cells induced by BMP9 via downregulation of BMP signaling pathway

doi: 10.3892/ijmm.2018.3526

Figure Lengend Snippet: miR-155 acts as an inhibitor of osteogenesis via suppressing BMP signaling during osteogenesis induced by bone morphogenetic protein 9 (BMP9) in C2C12 cells. (A) After treatment of C2C12 cells, the expression of p-Smad1/5/8 and runt-related transcription factor 2 (Runx2) were detected by western blotting. (B) The protein expression levels of osteocalcin (OCN) and OPN were measured by western blotting. β-actin was used as the internal controls, ** p<0.01 and *** p<0.001.

Article Snippet: Specific protein bands were detected with mouse anti-BMPR2 polyclonal antibody (1:200 dilution, cat. no. BM2821; BOSTER), rabbit anti-p-Smad1/5/8 monoclonal antibody (1:1,000 dilution, cat. no. sc-12353), rabbit anti-Smad1/5/8 monoclonal antibody (1:1,000 dilution, cat. no. sc-6031-R, lot no. D1911), rabbit anti-Runx2 monoclonal antibody (1:1,000 dilution, cat. no. sc-10758, lot no. A0810) (all from Santa Cruz Biotechnology, Inc., Santa Cruz, CA, USA), rabbit anti-osteocalcin (OCN) polyclonal antibody (1:200 dilution, cat. no. bs-4917R; Beijing Biosynthesis Biotechnology Co., Ltd.), rabbit anti-osteopontin (OPN) polyclonal antibody (1:500 dilution, cat. no. wl00691; Wanleibio, Shenyang, China), or anti-β-actin mouse monoclonal antibody (1:1,000 dilution; Zhongshan Golden Bridge Biotechnology Co., Ltd., Beijing, China) overnight at 4°C, and then washed with TBST three times, each time for 15 min, the membranes were incubated with goat anti-rabbit or goat anti-mouse horseradish peroxidase conjugated secondary antibody (1:5,000 dilution; Zhongshan Golden Bridge Biotechnology Co., Ltd.), for 1 h at 37°C.

Techniques: Expressing, Western Blot

Journal: International Journal of Molecular Medicine

Article Title: MicroRNA-155 inhibits the osteogenic differentiation of mesenchymal stem cells induced by BMP9 via downregulation of BMP signaling pathway

doi: 10.3892/ijmm.2018.3526

Figure Lengend Snippet: miR-155 acts as an inhibitor of osteogenesis via suppressing BMP signaling during the osteogenesis induced by bone morphogenetic protein 9 (BMP9) in mouse embryonic fibroblasts (MEF) cells. (A) After treatment of MEF cells, the expression of p-Smad1/5/8 and runt-related transcription factor 2 (Runx2) was detected by western blotting. (B) The protein expression levels of osteocalcin (OCN) and osteopontin (OPN) were measured by western blotting. β-actin was used as the internal controls, * p<0.05.

Article Snippet: Specific protein bands were detected with mouse anti-BMPR2 polyclonal antibody (1:200 dilution, cat. no. BM2821; BOSTER), rabbit anti-p-Smad1/5/8 monoclonal antibody (1:1,000 dilution, cat. no. sc-12353), rabbit anti-Smad1/5/8 monoclonal antibody (1:1,000 dilution, cat. no. sc-6031-R, lot no. D1911), rabbit anti-Runx2 monoclonal antibody (1:1,000 dilution, cat. no. sc-10758, lot no. A0810) (all from Santa Cruz Biotechnology, Inc., Santa Cruz, CA, USA), rabbit anti-osteocalcin (OCN) polyclonal antibody (1:200 dilution, cat. no. bs-4917R; Beijing Biosynthesis Biotechnology Co., Ltd.), rabbit anti-osteopontin (OPN) polyclonal antibody (1:500 dilution, cat. no. wl00691; Wanleibio, Shenyang, China), or anti-β-actin mouse monoclonal antibody (1:1,000 dilution; Zhongshan Golden Bridge Biotechnology Co., Ltd., Beijing, China) overnight at 4°C, and then washed with TBST three times, each time for 15 min, the membranes were incubated with goat anti-rabbit or goat anti-mouse horseradish peroxidase conjugated secondary antibody (1:5,000 dilution; Zhongshan Golden Bridge Biotechnology Co., Ltd.), for 1 h at 37°C.

Techniques: Expressing, Western Blot

Journal: International Journal of Molecular Medicine

Article Title: MicroRNA-155 inhibits the osteogenic differentiation of mesenchymal stem cells induced by BMP9 via downregulation of BMP signaling pathway

doi: 10.3892/ijmm.2018.3526

Figure Lengend Snippet: miR-155 directly targets runt-related transcription factor 2 (Runx2) and BMPR2. C2C12 cells and mouse embryonic fibroblasts (MEF) were transfected with NC, miR-155 or anti-miR-155, respectively. (A) Detected the expression of miR-155 by RT-qPCR. (B) Tested the expression of Runx2 and BMPR2 in C2C12 cells by RT-qPCR. (C) The expression of Runx2 and BMPR2 in C2C12 cells by western blotting. (D) The expression of Runx2 and BMPR2 in MEF cells by RT-qPCR. (E) The expression of Runx2 and BMPR2 in MEF cells by western blotting. (F) Putative binding sites for miR-155 in the 3′-untranslated region (3′-UTR) of Runx2 and BMPR2 as predicted by TargetScan and PicTar. Mutated nucleotides in mutant Runx2 3′-UTR and BMPR2 3′-UTR seed sequences are represented in blue. (G) Luciferase reporter assays in HEK-293 cells. Cells were co-transfected with WT-Runx2, or MT-Runx2, or WT-BMPR2 or MT-BMPR2 reporter construct or miR-155 overnight. Forty-eight hours post transfection luciferase activities were tested. Results were normalized to the value of cells that transfected pMIR-REPORT only. β-actin was used as the internal controls for western blotting, ** p<0.01 and *** p<0.001.

Article Snippet: Specific protein bands were detected with mouse anti-BMPR2 polyclonal antibody (1:200 dilution, cat. no. BM2821; BOSTER), rabbit anti-p-Smad1/5/8 monoclonal antibody (1:1,000 dilution, cat. no. sc-12353), rabbit anti-Smad1/5/8 monoclonal antibody (1:1,000 dilution, cat. no. sc-6031-R, lot no. D1911), rabbit anti-Runx2 monoclonal antibody (1:1,000 dilution, cat. no. sc-10758, lot no. A0810) (all from Santa Cruz Biotechnology, Inc., Santa Cruz, CA, USA), rabbit anti-osteocalcin (OCN) polyclonal antibody (1:200 dilution, cat. no. bs-4917R; Beijing Biosynthesis Biotechnology Co., Ltd.), rabbit anti-osteopontin (OPN) polyclonal antibody (1:500 dilution, cat. no. wl00691; Wanleibio, Shenyang, China), or anti-β-actin mouse monoclonal antibody (1:1,000 dilution; Zhongshan Golden Bridge Biotechnology Co., Ltd., Beijing, China) overnight at 4°C, and then washed with TBST three times, each time for 15 min, the membranes were incubated with goat anti-rabbit or goat anti-mouse horseradish peroxidase conjugated secondary antibody (1:5,000 dilution; Zhongshan Golden Bridge Biotechnology Co., Ltd.), for 1 h at 37°C.

Techniques: Transfection, Expressing, Quantitative RT-PCR, Western Blot, Binding Assay, Mutagenesis, Luciferase, Construct

Journal: International Journal of Molecular Medicine

Article Title: MicroRNA-155 inhibits the osteogenic differentiation of mesenchymal stem cells induced by BMP9 via downregulation of BMP signaling pathway

doi: 10.3892/ijmm.2018.3526

Figure Lengend Snippet: Interference of runt-related transcription factor 2 (Runx2) and BMPR2 in C2C12 cells and mouse embryonic fibroblasts (MEF), respectively. (A) The interference efficiency of si-Runx2 and shBMPR2 in C2C12 cells and mouse embryonic fibroblasts (MEF) after transfection by RT-qPCR. (B) The interference efficiency of si-Runx2 and shBMPR2 in C2C12 cells and MEF cells after transfection by western blotting. β-actin was used as the internal controls, *** p<0.001.

Article Snippet: Specific protein bands were detected with mouse anti-BMPR2 polyclonal antibody (1:200 dilution, cat. no. BM2821; BOSTER), rabbit anti-p-Smad1/5/8 monoclonal antibody (1:1,000 dilution, cat. no. sc-12353), rabbit anti-Smad1/5/8 monoclonal antibody (1:1,000 dilution, cat. no. sc-6031-R, lot no. D1911), rabbit anti-Runx2 monoclonal antibody (1:1,000 dilution, cat. no. sc-10758, lot no. A0810) (all from Santa Cruz Biotechnology, Inc., Santa Cruz, CA, USA), rabbit anti-osteocalcin (OCN) polyclonal antibody (1:200 dilution, cat. no. bs-4917R; Beijing Biosynthesis Biotechnology Co., Ltd.), rabbit anti-osteopontin (OPN) polyclonal antibody (1:500 dilution, cat. no. wl00691; Wanleibio, Shenyang, China), or anti-β-actin mouse monoclonal antibody (1:1,000 dilution; Zhongshan Golden Bridge Biotechnology Co., Ltd., Beijing, China) overnight at 4°C, and then washed with TBST three times, each time for 15 min, the membranes were incubated with goat anti-rabbit or goat anti-mouse horseradish peroxidase conjugated secondary antibody (1:5,000 dilution; Zhongshan Golden Bridge Biotechnology Co., Ltd.), for 1 h at 37°C.

Techniques: Transfection, Quantitative RT-PCR, Western Blot

Journal: International Journal of Molecular Medicine

Article Title: MicroRNA-155 inhibits the osteogenic differentiation of mesenchymal stem cells induced by BMP9 via downregulation of BMP signaling pathway

doi: 10.3892/ijmm.2018.3526

Figure Lengend Snippet: Knockdown of runt-related transcription factor 2 (Runx2) and BMPR2 separately reduced the effect of miR-155 in C2C12 cells. (A and B) C2C12 cells were co-transfected with RFP, or si-Runx2, or shNC, or shBMPR2, or miR-155, then induced osteogenic differentiation by bone morphogenetic protein 9 (BMP9) for 3 days to test the expression of Runx2, osterix (OSX), and induced for 7 days to test the expression of alkaline phosphatase (ALP), osteocalcin (OCN) used RT-qPCR. * p<0.05, ** p<0.01 and *** p<0.001.

Article Snippet: Specific protein bands were detected with mouse anti-BMPR2 polyclonal antibody (1:200 dilution, cat. no. BM2821; BOSTER), rabbit anti-p-Smad1/5/8 monoclonal antibody (1:1,000 dilution, cat. no. sc-12353), rabbit anti-Smad1/5/8 monoclonal antibody (1:1,000 dilution, cat. no. sc-6031-R, lot no. D1911), rabbit anti-Runx2 monoclonal antibody (1:1,000 dilution, cat. no. sc-10758, lot no. A0810) (all from Santa Cruz Biotechnology, Inc., Santa Cruz, CA, USA), rabbit anti-osteocalcin (OCN) polyclonal antibody (1:200 dilution, cat. no. bs-4917R; Beijing Biosynthesis Biotechnology Co., Ltd.), rabbit anti-osteopontin (OPN) polyclonal antibody (1:500 dilution, cat. no. wl00691; Wanleibio, Shenyang, China), or anti-β-actin mouse monoclonal antibody (1:1,000 dilution; Zhongshan Golden Bridge Biotechnology Co., Ltd., Beijing, China) overnight at 4°C, and then washed with TBST three times, each time for 15 min, the membranes were incubated with goat anti-rabbit or goat anti-mouse horseradish peroxidase conjugated secondary antibody (1:5,000 dilution; Zhongshan Golden Bridge Biotechnology Co., Ltd.), for 1 h at 37°C.

Techniques: Knockdown, Transfection, Expressing, Quantitative RT-PCR

Journal: International Journal of Molecular Medicine

Article Title: MicroRNA-155 inhibits the osteogenic differentiation of mesenchymal stem cells induced by BMP9 via downregulation of BMP signaling pathway

doi: 10.3892/ijmm.2018.3526

Figure Lengend Snippet: Knockdown runt-related transcription factor 2 (Runx2) and BMPR2 separately reduced the effect of miR-155 in mouse embryonic fibroblasts (MEF) cells. (A and B) MEF cells were co-transfected with RFP or si-Runx2 or shNC or shBMPR2 and miR-155, then induced osteogenic differentiation by BMP9 for 3 days to test the expression of Runx2, osterix (OSX), was induced for 7 days to test the expression of alkaline phosphatase (ALP) and osteocalcin (OCN) by RT-qPCR. * p<0.05, ** p<0.01 and *** p<0.001.

Article Snippet: Specific protein bands were detected with mouse anti-BMPR2 polyclonal antibody (1:200 dilution, cat. no. BM2821; BOSTER), rabbit anti-p-Smad1/5/8 monoclonal antibody (1:1,000 dilution, cat. no. sc-12353), rabbit anti-Smad1/5/8 monoclonal antibody (1:1,000 dilution, cat. no. sc-6031-R, lot no. D1911), rabbit anti-Runx2 monoclonal antibody (1:1,000 dilution, cat. no. sc-10758, lot no. A0810) (all from Santa Cruz Biotechnology, Inc., Santa Cruz, CA, USA), rabbit anti-osteocalcin (OCN) polyclonal antibody (1:200 dilution, cat. no. bs-4917R; Beijing Biosynthesis Biotechnology Co., Ltd.), rabbit anti-osteopontin (OPN) polyclonal antibody (1:500 dilution, cat. no. wl00691; Wanleibio, Shenyang, China), or anti-β-actin mouse monoclonal antibody (1:1,000 dilution; Zhongshan Golden Bridge Biotechnology Co., Ltd., Beijing, China) overnight at 4°C, and then washed with TBST three times, each time for 15 min, the membranes were incubated with goat anti-rabbit or goat anti-mouse horseradish peroxidase conjugated secondary antibody (1:5,000 dilution; Zhongshan Golden Bridge Biotechnology Co., Ltd.), for 1 h at 37°C.

Techniques: Knockdown, Transfection, Expressing, Quantitative RT-PCR

Journal: IEEE Open Journal of Nanotechnology

Article Title: Osteogenic Effect of Rabbit Periosteum-Derived Precursor Cells Co-Induced by Electric Stimulation and Adipose-Derived Stem Cells in a 3D Co-Culture System

doi: 10.1109/ojnano.2021.3131653

Figure Lengend Snippet: FIGURE 7. Double immunofluorescence staining showing protein expressions of the PDPCs after receiving ES treatment and co-culturing with ADSCs for 2 days. The ES treatment conditions were 0.7 v/cm, 80 kHz, and 3 h/day. The cell nuclei was stained by DPAI fluorescence (blue). The RUNX2 protein was stained by FITC fluorescence (green). The OPN protein was stained by iFlour 594 fluorescence (red).

Article Snippet: After washing, the cell-hydrogel construct was incubated with primary antibodies including diluted FITC-conjugated rabbit anti-RUNX2 polyclonal antibody (ARP36679; AVIVA SYSTEMS BIOSYSTEM, USA) and mouse anti-OPN monoclonal antibody (MBS555003; MyBioSource, USA) in PBS for 2 h at room temperature or overnight at 4 °C avoiding from the light.

Techniques: Staining

Journal: American journal of physiology. Heart and circulatory physiology

Article Title: Rapamycin Reduces Mineral Density and Promotes Beneficial Vascular Remodeling in a Murine Model of Severe Medial Arterial Calcification

doi: 10.1152/ajpheart.00530.2024

Figure Lengend Snippet: A. Graphic illustration of vehicle (DMSO) or 5 mg/kg rapamycin treatment for 25 days in Mgp +/+ and Mgp −/− mice (Created in BioRender. Behzadi, P. (2024) https://BioRender.com/r43g122 ). B. Representative microCT measurement of axial and cross sections of Mgp −/− mice aortas treated with vehicle or 5 mg/kg rapamycin, administered either once or three times a week for 25 days. Scale bar 0.2 mm. n=4 Mgp −/− 1x rapamycin, n=10 Mgp −/− 3x rapamycin, n=12 Mgp −/− vehicle C. Representative Von Kossa staining images of Mgp +/+ and Mgp −/− mice aortas treated with vehicle or rapamycin three times a week for 25 days. Scale bar 0.2 mm. n=10 Mgp −/− rapamycin, n=10 Mgp −/− vehicle, n=10 Mgp +/+ rapamycin, n=10 Mgp +/+ vehicle. D. Representative immunofluorescent images of Runx2 on the Mgp +/+ and Mgp −/− mice aortas treated with vehicle or rapamycin three times a week for 25 days. Scale bar 50 μm. n=10 Mgp −/− rapamycin, n=9 Mgp −/− vehicle, n=10 Mgp +/+ rapamycin, n=10 Mgp +/+ vehicle. Data are shown as means ± SD. P values were calculated using Kruskal-Wallis test (Figure B) and Two-way ANOVA test (Figures C, D).

Article Snippet: Tissues were incubated overnight at 4°C with antibodies for α-SMA (Abcam, ab5694, RRID: AB_2223021, 1:50), Ki67 (Abcam, ab15580, RRID: AB_443209, 1:250), LC3 (Abcam, ab192890, RRID: AB_2827794, 1:50), Mgp (Proteintech, 10734-I-AP, RRID: AB_2297660, 1:25), Myh11 (Invitrogen, MA5–4284, RRID: AB_2911986, 1:50), Runx2 (ABclonal, A2851, RRID: AB_2764676 1:100).

Techniques: Staining

Journal: Molecular Medicine Reports

Article Title: LINC01638 silencing inhibits cancer cell proliferation in colorectal adenocarcinoma through interaction with RUNX2

doi: 10.3892/mmr.2019.10191

Figure Lengend Snippet: Plasma RUNX2 mRNA levels are upregulated in colorectal adenocarcinoma patients and are positively correlated with the levels of lncRNA LINC01638. (A) Plasma RUNX2 mRNA levels were upregulated in colorectal adenocarcinoma patients compared to healthy controls (*P<0.05). Plasma levels of RUNX2 mRNA and lncRNA LINC01638 were positively correlated in (B) colorectal adenocarcinoma patients but not in (C) healthy controls.

Article Snippet: Antibodies used in this study included primary antibodies of rabbit anti-human RUNX2 (1:1,500; cat. no. ab23981; Abcam, Shanghai, China) and (1:1,500; cat. no. ab9485; Abcam) and secondary antibody goat anti-rabbit IgG-HRP (1:1,200; cat. no. MBS435036; MyBioSource, San Diego, CA, USA).

Techniques: Clinical Proteomics

Journal: Molecular Medicine Reports

Article Title: LINC01638 silencing inhibits cancer cell proliferation in colorectal adenocarcinoma through interaction with RUNX2

doi: 10.3892/mmr.2019.10191

Figure Lengend Snippet: LINC01638 shRNA silencing mediates downreguatin of RUNX2 in cells of WiDr and HT-29 human colorectal adenocarcinoma cell lines. (A) LINC01638 shRNA silencing led to significantly downregulated expression of RUNX2 protein in cells of WiDr and HT-29 human colorectal adenocarcinoma cell lines, while (B) RUNX2 overexpression failed to significanly affect the expression of LINC01638 (*P<0.05).

Article Snippet: Antibodies used in this study included primary antibodies of rabbit anti-human RUNX2 (1:1,500; cat. no. ab23981; Abcam, Shanghai, China) and (1:1,500; cat. no. ab9485; Abcam) and secondary antibody goat anti-rabbit IgG-HRP (1:1,200; cat. no. MBS435036; MyBioSource, San Diego, CA, USA).

Techniques: shRNA, Expressing, Over Expression

Journal: Molecular Medicine Reports

Article Title: LINC01638 silencing inhibits cancer cell proliferation in colorectal adenocarcinoma through interaction with RUNX2

doi: 10.3892/mmr.2019.10191

Figure Lengend Snippet: LINC01638 shRNA inhibits cancer cell proliferation possibly through RUNX2. LINC01638 shRNA silencing (shRNA) significantly inhibited, while RUNX2 overexpression (RUNX2) significantly promoted the proliferation of cells of both (A) WiDr and (B) HT-29 human colorectal adenocarcinoma cell lines. In addition, RUNX2 overexpression partially reversed the advese effect of LINC01638 shRNA silencing on cancer cell proliferation (*P<0.05).

Article Snippet: Antibodies used in this study included primary antibodies of rabbit anti-human RUNX2 (1:1,500; cat. no. ab23981; Abcam, Shanghai, China) and (1:1,500; cat. no. ab9485; Abcam) and secondary antibody goat anti-rabbit IgG-HRP (1:1,200; cat. no. MBS435036; MyBioSource, San Diego, CA, USA).

Techniques: shRNA, Over Expression