Journal: Respiratory Research

Article Title: The role and mechanism of thrombospondin-4 in pulmonary arterial hypertension associated with congenital heart disease

doi: 10.1186/s12931-024-02932-w

Figure Lengend Snippet: Circulating level of THBS4 in patients with PAH-CHD. The resultant data are represented as mean ± SD. * P < 0.05

Article Snippet: The PVDF membrane was blocked for nonspecific binding with blocking buffer and incubated with primary antibodies against THBS4 (Abcam, USA, 1:1000), GAPDH (Proteintech, China, 1:20000), α-SMA (Proteintech, China, 1:20000), MYH11 (SANTA CRUZ, America, 1:200), PCNA (SANTA CRUZ, America, 1:200), N-cad (SANTA CRUZ, America, 1:200), BAX (Abmart, China, 1:1000), AKT (CST, America,1:1000), PI3K (CST, America, 1:1000), p-AKT (Abmart, China, 1:1000), and p-PI3K (Abmart, China, 1:1000) overnight at 4 ℃.

Techniques:

Journal: Respiratory Research

Article Title: The role and mechanism of thrombospondin-4 in pulmonary arterial hypertension associated with congenital heart disease

doi: 10.1186/s12931-024-02932-w

Figure Lengend Snippet: Expression of THBS4 in PAH rats induced by MCT plus aorto-caval shunt. (A) THBS4 expression in GSE149899. qRT-PCR (B) and Western blot (C) showed the time course of THBS4 expression in PAH rats induced by MCT plus aorto-caval shunt. (D) Double immunofluorescence staining of THBS4 with α-SMA or VWF. Scare bars: 50 μm. CON: the control group; SHAM: the sham group; MS: PAH rats induced by MCT plus aorto-caval shunt (MS) from 1 week (1w) to 4 weeks (4w). The resultant data are represented as mean ± SD. * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001

Article Snippet: The PVDF membrane was blocked for nonspecific binding with blocking buffer and incubated with primary antibodies against THBS4 (Abcam, USA, 1:1000), GAPDH (Proteintech, China, 1:20000), α-SMA (Proteintech, China, 1:20000), MYH11 (SANTA CRUZ, America, 1:200), PCNA (SANTA CRUZ, America, 1:200), N-cad (SANTA CRUZ, America, 1:200), BAX (Abmart, China, 1:1000), AKT (CST, America,1:1000), PI3K (CST, America, 1:1000), p-AKT (Abmart, China, 1:1000), and p-PI3K (Abmart, China, 1:1000) overnight at 4 ℃.

Techniques: Expressing, Quantitative RT-PCR, Western Blot, Double Immunofluorescence Staining, Control

Journal: Respiratory Research

Article Title: The role and mechanism of thrombospondin-4 in pulmonary arterial hypertension associated with congenital heart disease

doi: 10.1186/s12931-024-02932-w

Figure Lengend Snippet: qRT-PCR (A) and Western blot (B) showed the transfection efficiency of THBS4 in PASMCs. (C) The influence of THBS4 on the transformation of phenotype of PASMCs. The resultant data are represented as mean ± SD. *: si-THBS4 vs.si-CON; #: pEX3-THBS4 vs. pEX3-CON. * P < 0.05; *** P < 0.001; # P < 0.05; ## P < 0.01; ### P < 0.001

Article Snippet: The PVDF membrane was blocked for nonspecific binding with blocking buffer and incubated with primary antibodies against THBS4 (Abcam, USA, 1:1000), GAPDH (Proteintech, China, 1:20000), α-SMA (Proteintech, China, 1:20000), MYH11 (SANTA CRUZ, America, 1:200), PCNA (SANTA CRUZ, America, 1:200), N-cad (SANTA CRUZ, America, 1:200), BAX (Abmart, China, 1:1000), AKT (CST, America,1:1000), PI3K (CST, America, 1:1000), p-AKT (Abmart, China, 1:1000), and p-PI3K (Abmart, China, 1:1000) overnight at 4 ℃.

Techniques: Quantitative RT-PCR, Western Blot, Transfection, Transformation Assay

Journal: Respiratory Research

Article Title: The role and mechanism of thrombospondin-4 in pulmonary arterial hypertension associated with congenital heart disease

doi: 10.1186/s12931-024-02932-w

Figure Lengend Snippet: Effect of THBS4 on the proliferation, apoptosis, and migration of PASMCs. (A) Effect of THBS4 on proliferation of PASMCs, as determined by the EdU assay. Scare bars, 100 μm. (B) Effect of THBS4 on apoptosis of PASMCs, as determined by the Annexin-V flow cytometry. (C) Effect of THBS4 on migration of PASMCs, as determined by the Wound healing assay. Scare bars, 200 μm. (D) Effect of THBS4 on migration of PASMCs, as determined by the Transwell assay. Scare bars, 100 μm. The resultant data are represented as mean ± SD. *: si-THBS4 vs.si-CON; #: pEX3-THBS4 vs. pEX3-CON. * P < 0.05; ** P < 0.01; **** P < 0.0001; # P < 0.05; ## P < 0.01

Article Snippet: The PVDF membrane was blocked for nonspecific binding with blocking buffer and incubated with primary antibodies against THBS4 (Abcam, USA, 1:1000), GAPDH (Proteintech, China, 1:20000), α-SMA (Proteintech, China, 1:20000), MYH11 (SANTA CRUZ, America, 1:200), PCNA (SANTA CRUZ, America, 1:200), N-cad (SANTA CRUZ, America, 1:200), BAX (Abmart, China, 1:1000), AKT (CST, America,1:1000), PI3K (CST, America, 1:1000), p-AKT (Abmart, China, 1:1000), and p-PI3K (Abmart, China, 1:1000) overnight at 4 ℃.

Techniques: Migration, EdU Assay, Flow Cytometry, Wound Healing Assay, Transwell Assay

Journal: Respiratory Research

Article Title: The role and mechanism of thrombospondin-4 in pulmonary arterial hypertension associated with congenital heart disease

doi: 10.1186/s12931-024-02932-w

Figure Lengend Snippet: Phosphorylation level of PI3K/AKT in PASMCs after inhibition and overexpression of THBS4 in PASMCs. The resultant data are represented as mean ± SD. *: si-THBS4 vs. si-CON; #: pEX3-THBS4 vs. pEX3-CON. * P < 0.5; # P < 0.5

Article Snippet: The PVDF membrane was blocked for nonspecific binding with blocking buffer and incubated with primary antibodies against THBS4 (Abcam, USA, 1:1000), GAPDH (Proteintech, China, 1:20000), α-SMA (Proteintech, China, 1:20000), MYH11 (SANTA CRUZ, America, 1:200), PCNA (SANTA CRUZ, America, 1:200), N-cad (SANTA CRUZ, America, 1:200), BAX (Abmart, China, 1:1000), AKT (CST, America,1:1000), PI3K (CST, America, 1:1000), p-AKT (Abmart, China, 1:1000), and p-PI3K (Abmart, China, 1:1000) overnight at 4 ℃.

Techniques: Inhibition, Over Expression

Journal: Respiratory Research

Article Title: The role and mechanism of thrombospondin-4 in pulmonary arterial hypertension associated with congenital heart disease

doi: 10.1186/s12931-024-02932-w

Figure Lengend Snippet: Effect of THBS4 suppression on pulmonary vascular remodeling in PAH rats induced by MCT plus aorto-caval shunt. (A) The AAV transfection efficiency was determined by detecting green fluorescence protein which is encoded by AAV in lung tissues. Scare bars, 100 μm. (B) THBS4 suppression efficiency determined by immunofluorescence staining of THBS4. Scare bars, 50 μm. (C) Hematoxylin and Eosin staining of lung tissues. Scare bars,100 μm. (D) Immunofluorescence staining of α-SMA in lung tissues. Scare bars,100 μm. The percentage of non-muscularized (E) , partially-muscularized (F), and fully-muscularized pulmonary arterioles (G) . (H) The percentage of wall thickness. (I) The percentage of wall area. (G) Cross-sectional area of the right ventricular cardiomyocyte. Scare bars,100 μm. The resultant data are represented as mean ± SD. CON: control group; SHAM: sham group; MS-AAV-CON: administration of control AAV + MCT plus aorto-caval shunt induced PAH rats; MS-AAV-siTHBS4: administration of THBS4 suppression AAV + MCT plus aorto-caval shunt induced PAH rats. PA: pulmonary arteriole. * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001

Article Snippet: The PVDF membrane was blocked for nonspecific binding with blocking buffer and incubated with primary antibodies against THBS4 (Abcam, USA, 1:1000), GAPDH (Proteintech, China, 1:20000), α-SMA (Proteintech, China, 1:20000), MYH11 (SANTA CRUZ, America, 1:200), PCNA (SANTA CRUZ, America, 1:200), N-cad (SANTA CRUZ, America, 1:200), BAX (Abmart, China, 1:1000), AKT (CST, America,1:1000), PI3K (CST, America, 1:1000), p-AKT (Abmart, China, 1:1000), and p-PI3K (Abmart, China, 1:1000) overnight at 4 ℃.

Techniques: Transfection, Fluorescence, Immunofluorescence, Staining, Control

Journal: Aging (Albany NY)

Article Title: THBS4/integrin α2 axis mediates BM-MSCs to promote angiogenesis in gastric cancer associated with chronic Helicobacter pylori infection

doi: 10.18632/aging.203334

Figure Lengend Snippet: BM-MSC engraftment leads to THBS4 overexpression during H. pylori infection. ( A ) Volcano plot presentation of differentially expressed genes of gastric tissues in H. pylori versus H. pylori + MSCs mice. ( B ) Enrichment analysis of differential genes. ( C ) RT-PCR verified the expression of differential genes associated with angiogenesis. THBS4 was the most highly expressed gene. n= 10 mice in each group. ( D , E ) Western blot analysis showed that BM-MSC transplantation led to THBS4 overexpression. n= 3 mice in each group. ( F , G ) IHC ( F ) and IF ( G ) of THBS4 in H. pylori + MSCs or H. pylori mice. n= 3 mice per group Cell nuclei, DAPI. THBS4, FITC. Magnification: × 200 and × 400. ( H ) Quantification of F. ( I , J ) Western blot analysis of THBS4 revealed that H. pylori pretreatment increased the protein expression of THBS4 in BM-MSCs. n= 3 wells per group.

Article Snippet: After dewaxing and hydration, the sections were incubated overnight with antibodies against human THBS4 (YT4646, ImmunoWay), human PECAM-1 (ab9498, Abcam), mouse CD31 (AF3628-SP, R&D Systems), mouse NG2 (sc-5389, Santa Cruz), mouse THBS4 (AF7860-SP R&D Systems), and mouse Ki-67 (ab1667, Abcam) and identified with Alexa 488, Alexa 594 or Cy3 secondary antibodies or HRP-conjugated secondary antibodies the next day.

Techniques: Over Expression, Infection, Reverse Transcription Polymerase Chain Reaction, Expressing, Western Blot, Transplantation Assay

Journal: Aging (Albany NY)

Article Title: THBS4/integrin α2 axis mediates BM-MSCs to promote angiogenesis in gastric cancer associated with chronic Helicobacter pylori infection

doi: 10.18632/aging.203334

Figure Lengend Snippet: Overexpression of THBS4 is correlated with vessel density in human GC tissues and predicts a poor prognosis. ( A ) The expression of THBS4 in a pan-cancer dataset. The red and green labels at the top represent high and low expression in certain tumor types, respectively. ( B ) Volcano plot revealed upregulation of THBS4 mRNA expression in GC compared with normal tissue. ( C , D ) The overall survival and disease-free survival analyses of THBS4 were plotted for patients with GC in TCGA. ( E ) Representative IHC staining with THBS4 antibody in GC and adjacent nontumor tissues. n= 5 samples per group. ( F ) Quantification of E. ( G ) Representative IHC staining with PECAM1 antibody in GC and adjacent nontumor tissues. n= 5 samples per group. ( H ) Quantification of G. ( I ) Correlation analysis of THBS4 and vessel density showed that THBS4 was positively correlated with vessel density in GC. ( J ) Validation of the positive correlation between THBS4 and PECAM1 in GEPIA.

Article Snippet: After dewaxing and hydration, the sections were incubated overnight with antibodies against human THBS4 (YT4646, ImmunoWay), human PECAM-1 (ab9498, Abcam), mouse CD31 (AF3628-SP, R&D Systems), mouse NG2 (sc-5389, Santa Cruz), mouse THBS4 (AF7860-SP R&D Systems), and mouse Ki-67 (ab1667, Abcam) and identified with Alexa 488, Alexa 594 or Cy3 secondary antibodies or HRP-conjugated secondary antibodies the next day.

Techniques: Over Expression, Expressing, Immunohistochemistry, Biomarker Discovery

Journal: Aging (Albany NY)

Article Title: THBS4/integrin α2 axis mediates BM-MSCs to promote angiogenesis in gastric cancer associated with chronic Helicobacter pylori infection

doi: 10.18632/aging.203334

Figure Lengend Snippet: THBS4 mediates BM-MSCs to promote the migration and tube formation of HUVECs in vitro . ( A ) Transfection of recombinant lentiviral vectors into BM-MSCs. ( B – E ) qRT-PCR and Western blot analysis of THBS4 expression in BM-MSCs of both experiments transfected as indicated. n= 3 wells per group. ( F , H ) Transwell assays and quantification of migrated cells showed that THBS4 knockdown in BM-MSCs inhibited their ability to promote HUVEC migration. n= 3 wells in each group. ( G , I ) Knockdown of THBS4 in BM-MSCs alleviated its ability to promote HUVEC tube formation. n= 5 wells per group.

Article Snippet: After dewaxing and hydration, the sections were incubated overnight with antibodies against human THBS4 (YT4646, ImmunoWay), human PECAM-1 (ab9498, Abcam), mouse CD31 (AF3628-SP, R&D Systems), mouse NG2 (sc-5389, Santa Cruz), mouse THBS4 (AF7860-SP R&D Systems), and mouse Ki-67 (ab1667, Abcam) and identified with Alexa 488, Alexa 594 or Cy3 secondary antibodies or HRP-conjugated secondary antibodies the next day.

Techniques: Migration, In Vitro, Transfection, Recombinant, Quantitative RT-PCR, Western Blot, Expressing, Knockdown

Journal: Aging (Albany NY)

Article Title: THBS4/integrin α2 axis mediates BM-MSCs to promote angiogenesis in gastric cancer associated with chronic Helicobacter pylori infection

doi: 10.18632/aging.203334

Figure Lengend Snippet: THBS4 mediates BM-MSC-induced promotion of GC angiogenesis in vivo . ( A , B ) Four-week-old nude male mice were injected with either SGC cells only or SGC and BM-MSCs transfected with recombinant lentiviral vectors. Three weeks following injection, the mice were euthanized, and the tumors were analyzed. n= 3 mice in control group and 5 mice per group in other groups. ( C ) Growth curve of tumors described in A. ( D ) Tumor weight of injected tumors at end-point. ( E ) H&E staining and Ki-67 and CD31 immunostaining of tumors described in A. ( F , G ) Quantification of staining presented in E performed with Image Plus. ( H , I ) Following AngioSense 750EX injection, tumor vascular permeability fluorescence imaging in vivo was performed to detect the neovascularization density of tumors in A. ( J , K ) Representative images and quantification of the CAM assay. n= 5 eggs in each group. ( L, M ) Western blot analysis of THBS4 in gastric tissues after NC BM-MSC or THBS4-knockdown BM-MSC transplantation. n= 3 mice in each group. ( N ) IF of THBS4 in gastric tissues after NC BM-MSC or THBS4-knockdown BM-MSC transplantation. ( O ) IF of CD31 and NG2 in gastric tissues in NC BM-MSC-transplanted mice and THBS4-knockdown BM-MSC-transplanted mice. n= 10 mice in each group.

Article Snippet: After dewaxing and hydration, the sections were incubated overnight with antibodies against human THBS4 (YT4646, ImmunoWay), human PECAM-1 (ab9498, Abcam), mouse CD31 (AF3628-SP, R&D Systems), mouse NG2 (sc-5389, Santa Cruz), mouse THBS4 (AF7860-SP R&D Systems), and mouse Ki-67 (ab1667, Abcam) and identified with Alexa 488, Alexa 594 or Cy3 secondary antibodies or HRP-conjugated secondary antibodies the next day.

Techniques: In Vivo, Injection, Transfection, Recombinant, Control, Staining, Immunostaining, Permeability, Fluorescence, Imaging, Chick Chorioallantoic Membrane Assay, Western Blot, Knockdown, Transplantation Assay

Journal: Aging (Albany NY)

Article Title: THBS4/integrin α2 axis mediates BM-MSCs to promote angiogenesis in gastric cancer associated with chronic Helicobacter pylori infection

doi: 10.18632/aging.203334

Figure Lengend Snippet: Integrin α2 mediates the effect of paracrine THBS4 signaling on the migration and tube formation of HUVECs. ( A – C ) Western blotting and IF showed that integrin α2 in HUVECs was significantly increased by rTHBS4. n= 3 wells per group. ( D ) Representative images of the migration assay after rTHBS4, rTHBS4 + anti- α2 or anti- α2 treatments in HUVECs. n= three independent experiments. ( E ) Representative images of the tube formation assay after rTHBS4, rTHBS4 + anti-α2 or anti-α2 treatments in HUVECs. n= 3 wells in each group. ( F ) Quantification of D. ( G ) Quantification of E. ( H ) CCK-8 assay to detect HUVEC proliferation after rTHBS4, rTHBS4 + anti-α2 or anti-α2 treatments. n= three independent experiments.

Article Snippet: After dewaxing and hydration, the sections were incubated overnight with antibodies against human THBS4 (YT4646, ImmunoWay), human PECAM-1 (ab9498, Abcam), mouse CD31 (AF3628-SP, R&D Systems), mouse NG2 (sc-5389, Santa Cruz), mouse THBS4 (AF7860-SP R&D Systems), and mouse Ki-67 (ab1667, Abcam) and identified with Alexa 488, Alexa 594 or Cy3 secondary antibodies or HRP-conjugated secondary antibodies the next day.

Techniques: Migration, Western Blot, Tube Formation Assay, CCK-8 Assay

Journal: Aging (Albany NY)

Article Title: THBS4/integrin α2 axis mediates BM-MSCs to promote angiogenesis in gastric cancer associated with chronic Helicobacter pylori infection

doi: 10.18632/aging.203334

Figure Lengend Snippet: Activation of AKT by THBS4/integrin α2 axis in HUVECs. ( A , B ) rTHBS4 induced the phosphorylation of AKT in HUVECs. n= three independent experiments. The HUVECs were treated with and without rTHBS4 for 1h and the whole cell lysates were analyzed by WB for levels of downstream kinases. ( C , D ) p-AKT activation occurred in a time-dependent manner in rTHBS4-treated cells. n= three independent experiments. ( E , F ) The HUVECs were treated with rTHBS4 for 1h in the presence of integrin α2-specific antibody or PI3K inhibitor LY294002. Western blot analysis showed that rTHBS4 increased Akt phosphorylation in HUVECs, while this effect was alleviated by blocking integrin α2 or inhibiting Akt. n= three independent experiments. ( G ) Transwell migration assay to investigate the effect of PI3K inhibition and integrin α2 blockade on HUVEC migration after treatment with rTHBS4. n= 3 wells per group. ( H ) In vitro tube formation to investigate the effect of PI3K inhibition and blocking integrin α2 on HUVEC tube formation. n= 3 wells per group. ( I ) Quantification of G. ( J ) Quantification of H. ( K , L ) The CM from BM-MSCs stimulated by H. Pylori was applied to HUVECs in the presence or absence of integrin α2-specific antibody or PI3K inhibitor LY294002 for Transwell migration assay and tube formation assay. n= 3 wells per group. ( M ) Quantification of K. ( N ) Quantification of L.

Article Snippet: After dewaxing and hydration, the sections were incubated overnight with antibodies against human THBS4 (YT4646, ImmunoWay), human PECAM-1 (ab9498, Abcam), mouse CD31 (AF3628-SP, R&D Systems), mouse NG2 (sc-5389, Santa Cruz), mouse THBS4 (AF7860-SP R&D Systems), and mouse Ki-67 (ab1667, Abcam) and identified with Alexa 488, Alexa 594 or Cy3 secondary antibodies or HRP-conjugated secondary antibodies the next day.

Techniques: Activation Assay, Phospho-proteomics, Western Blot, Blocking Assay, Transwell Migration Assay, Inhibition, Migration, In Vitro, Tube Formation Assay

Journal: Frontiers in Pharmacology

Article Title: The Attenuating Effect of Low-Intensity Pulsed Ultrasound on Hypoxia-Induced Rat Chondrocyte Damage in TMJ Osteoarthritis Based on TMT Labeling Quantitative Proteomic Analysis

doi: 10.3389/fphar.2021.752734

Figure Lengend Snippet: Primer sequences used for RT-qPCR.

Article Snippet: The primary antibodies included rabbit polyclonal anti-collagen Ⅱ (1:3,000, ab34712, Abcam), rabbit monoclonal anti-aggrecan (1:1,000, A11691, ABclonal), rabbit polyclonal anti-MMP3 (1:1,000, ab52915, Abcam), rabbit polyclonal anti-TIMP1 (1:1,000, ab61224, Abcam), rabbit polyclonal anti-VEGFA (1:1,000, A12303, ABclonal), rabbit polyclonal anti-THBS4 (1:1,000, ab263898, Abcam), rabbit monoclonal anti-THBS1 (1:1,000, ab22928-10, Abcam), rabbit polyclonal anti-IL1RL1 (1:1,000, ab228543, Abcam), rabbit polyclonal anti-GLUL (1:1,000, ab176562, Abcam), and anti-β-actin (1:100,000, AC026, ABclonal) antibodies.

Techniques: Sequencing

Journal: Frontiers in Pharmacology

Article Title: The Attenuating Effect of Low-Intensity Pulsed Ultrasound on Hypoxia-Induced Rat Chondrocyte Damage in TMJ Osteoarthritis Based on TMT Labeling Quantitative Proteomic Analysis

doi: 10.3389/fphar.2021.752734

Figure Lengend Snippet: Top 5 DEPs identified from the L/N, LP/L and LP/N comparisons.

Article Snippet: The primary antibodies included rabbit polyclonal anti-collagen Ⅱ (1:3,000, ab34712, Abcam), rabbit monoclonal anti-aggrecan (1:1,000, A11691, ABclonal), rabbit polyclonal anti-MMP3 (1:1,000, ab52915, Abcam), rabbit polyclonal anti-TIMP1 (1:1,000, ab61224, Abcam), rabbit polyclonal anti-VEGFA (1:1,000, A12303, ABclonal), rabbit polyclonal anti-THBS4 (1:1,000, ab263898, Abcam), rabbit monoclonal anti-THBS1 (1:1,000, ab22928-10, Abcam), rabbit polyclonal anti-IL1RL1 (1:1,000, ab228543, Abcam), rabbit polyclonal anti-GLUL (1:1,000, ab176562, Abcam), and anti-β-actin (1:100,000, AC026, ABclonal) antibodies.

Techniques:

Journal: Frontiers in Pharmacology

Article Title: The Attenuating Effect of Low-Intensity Pulsed Ultrasound on Hypoxia-Induced Rat Chondrocyte Damage in TMJ Osteoarthritis Based on TMT Labeling Quantitative Proteomic Analysis

doi: 10.3389/fphar.2021.752734

Figure Lengend Snippet: Validation of DEPs identified from the comparisons of the N, L and LP groups. (A) Western blot analysis of proteins including THBS4, THBS1, IL1RL1, GLUL and TIMP1. With the exception of the IL1RL1 band, all bands on the Western blot were consistent with the bioinformatic analysis results. RT-qPCR analysis of the following genes: THBS4 (B) , THBS1 (C) , IL1RL1 (D) , GLUL (E) and TIMP1 (F) . The mRNA levels of Thbs4 , Thbs1 , IL1RL1 , and Glul were consistent with the trends observed in the bioinformatic analyses. The data are shown as the mean ± SD values; * p < 0.05, ** p < 0.01, and *** p < 0.001.

Article Snippet: The primary antibodies included rabbit polyclonal anti-collagen Ⅱ (1:3,000, ab34712, Abcam), rabbit monoclonal anti-aggrecan (1:1,000, A11691, ABclonal), rabbit polyclonal anti-MMP3 (1:1,000, ab52915, Abcam), rabbit polyclonal anti-TIMP1 (1:1,000, ab61224, Abcam), rabbit polyclonal anti-VEGFA (1:1,000, A12303, ABclonal), rabbit polyclonal anti-THBS4 (1:1,000, ab263898, Abcam), rabbit monoclonal anti-THBS1 (1:1,000, ab22928-10, Abcam), rabbit polyclonal anti-IL1RL1 (1:1,000, ab228543, Abcam), rabbit polyclonal anti-GLUL (1:1,000, ab176562, Abcam), and anti-β-actin (1:100,000, AC026, ABclonal) antibodies.

Techniques: Western Blot, Quantitative RT-PCR

Journal: Molecular Medicine Reports

Article Title: Reciprocal regulation of long noncoding RNAs THBS4-003 and THBS4 control migration and invasion in prostate cancer cell lines

doi: 10.3892/mmr.2016.5443

Figure Lengend Snippet: Expression of lncRNA-THBS4-003 in PCa and adjacent non-tumor tissues. (A) A microarray containing 8,277 lncRNA probes and 32,207 mRNA probes was used to identify dysregulated mRNAs in three patients with PCa. Of these, 354 mRNAs were significantly upregulated and 350 were significantly downregulated (P<0.05; FC>2; red = high expression, green = low expression). (B) Higher expression levels of THBS4 were found in PCa tissues, compared with adjacent non-tumor tissues. (C) Protein expression levels of THBS4 in three matched non-tumor/tumor tissues were detected using Western blot analysis. (D) Quantification of western blot analysis. Data are presented as the mean ± standard deviation. * P<0.05. β-actin was used as an internal control. PCa prostate cancer; THBS4, thrombospondin 4; N, non-tumor; T, tumor.

Article Snippet: The sequences of the siRNAs used in the present study were as follows: 5′-GGCAACAGCUACAGUACAATT-3′ and 5′-UUGUACUGUAGCUGUUGCCTT-3′ for si-lncRNA-THBS4-003, 5′-GGCAGUUCUUGGGUCAAAUTT-3′ and 5′-AUUUGACCCAAGAACUGCCTT-3′ for si-THBS4, and 5′-UUCUCCGAACGUGUCACGUTT-3′ and 5′-ACGUGACACGUUCGGAGAATT-3′ for the control siRNA (Invitrogen; Thermo Fisher Scientific, Inc.).

Techniques: Expressing, Microarray, Western Blot, Standard Deviation

Journal: Molecular Medicine Reports

Article Title: Reciprocal regulation of long noncoding RNAs THBS4-003 and THBS4 control migration and invasion in prostate cancer cell lines

doi: 10.3892/mmr.2016.5443

Figure Lengend Snippet: Expression of lncRNA-THBS4-003 in PCa and adjacent non-tumor tissues. (A) Expression levels of lncRNA-THBS4-003 were higher in the PCa tissues, compared with the adjacent non-tumor tissues. (B) Expression of lncRNA-THBS4-003 in patients with different Gleason scores. β-actin was used as a loading control. Data are presented as the mean ± standard error of the mean. * P<0.05 (determined using Student's t -test). (C) Expression levels of THBS4 and lncRNA-THBS4-003 were positively correlated in PCa tissues (P<0.0001). PCa, prostate cancer; lncRNA, long noncoding RNA; THBS4, thrombospondin 4.

Article Snippet: The sequences of the siRNAs used in the present study were as follows: 5′-GGCAACAGCUACAGUACAATT-3′ and 5′-UUGUACUGUAGCUGUUGCCTT-3′ for si-lncRNA-THBS4-003, 5′-GGCAGUUCUUGGGUCAAAUTT-3′ and 5′-AUUUGACCCAAGAACUGCCTT-3′ for si-THBS4, and 5′-UUCUCCGAACGUGUCACGUTT-3′ and 5′-ACGUGACACGUUCGGAGAATT-3′ for the control siRNA (Invitrogen; Thermo Fisher Scientific, Inc.).

Techniques: Expressing

Journal: Molecular Medicine Reports

Article Title: Reciprocal regulation of long noncoding RNAs THBS4-003 and THBS4 control migration and invasion in prostate cancer cell lines

doi: 10.3892/mmr.2016.5443

Figure Lengend Snippet: Effects of THBS4 knockdown on PCa cell lines. (A) Expression levels of THBS4 mRNA and lncRNA-THBS4-003 following transfection with mock, nc or si-THBS4. β-actin was used as a loading control. (B) Expression levels of THBS4, P38 and MMP-9 following transfection with mock, nc or si-THBS4. THBS4 knockdown inhibited the cell (C) migration and (D) invasion in vitro . Data are presented as the mean ± standard error of the mean of at least three independent experiments. * P<0.05. Original magnification ×200. PCa, prostate cancer; lncRNA, long noncoding RNA; THBS4, thrombospondin 4; si-THBS4, small interfering THBS4; MMP-9, matrix metallorpoteinase-9; nc, negative control.

Article Snippet: The sequences of the siRNAs used in the present study were as follows: 5′-GGCAACAGCUACAGUACAATT-3′ and 5′-UUGUACUGUAGCUGUUGCCTT-3′ for si-lncRNA-THBS4-003, 5′-GGCAGUUCUUGGGUCAAAUTT-3′ and 5′-AUUUGACCCAAGAACUGCCTT-3′ for si-THBS4, and 5′-UUCUCCGAACGUGUCACGUTT-3′ and 5′-ACGUGACACGUUCGGAGAATT-3′ for the control siRNA (Invitrogen; Thermo Fisher Scientific, Inc.).

Techniques: Expressing, Transfection, Migration, In Vitro, Negative Control

Journal: Molecular Medicine Reports

Article Title: Reciprocal regulation of long noncoding RNAs THBS4-003 and THBS4 control migration and invasion in prostate cancer cell lines

doi: 10.3892/mmr.2016.5443

Figure Lengend Snippet: Knockdown of lncRNA-THBS4-003 inhibits PCa cell line migration and invasion in vitro . (A) Expression levels of lncRNA-THBS4-003 in transfected PC-3 cells were measured using reverse transcription-quantitative polymerase chain reaction analysis. β-actin was used as a loading control (determined using Student's t -test). lncRNA-THBS4-003 knockdown inhibited cell (B) migration and (C) invasion in vitro . Data are presented as the mean ± standard error of the mean of at least three independent experiments. * P<0.05. Original magnification ×200. PCa, prostate cancer; lncRNA, long noncoding RNA; THBS4, thrombospondin 4; si-THBS4, small interfering-THBS4, nc, negative control.

Article Snippet: The sequences of the siRNAs used in the present study were as follows: 5′-GGCAACAGCUACAGUACAATT-3′ and 5′-UUGUACUGUAGCUGUUGCCTT-3′ for si-lncRNA-THBS4-003, 5′-GGCAGUUCUUGGGUCAAAUTT-3′ and 5′-AUUUGACCCAAGAACUGCCTT-3′ for si-THBS4, and 5′-UUCUCCGAACGUGUCACGUTT-3′ and 5′-ACGUGACACGUUCGGAGAATT-3′ for the control siRNA (Invitrogen; Thermo Fisher Scientific, Inc.).

Techniques: Migration, In Vitro, Expressing, Transfection, Real-time Polymerase Chain Reaction, Negative Control

Journal: Molecular Medicine Reports

Article Title: Reciprocal regulation of long noncoding RNAs THBS4-003 and THBS4 control migration and invasion in prostate cancer cell lines

doi: 10.3892/mmr.2016.5443

Figure Lengend Snippet: Expression levels of THBS4, P38 and MMP-9 are decreased following transfection with si-lncRNA-THBS4-003. * P<0.05. lncRNA, long noncoding RNA; THBS4, thrombospondin 4; si-THBS4, small interfering-THBS4; MMP-9, matrix metallorpoteinase-9; nc, negative control.

Article Snippet: The sequences of the siRNAs used in the present study were as follows: 5′-GGCAACAGCUACAGUACAATT-3′ and 5′-UUGUACUGUAGCUGUUGCCTT-3′ for si-lncRNA-THBS4-003, 5′-GGCAGUUCUUGGGUCAAAUTT-3′ and 5′-AUUUGACCCAAGAACUGCCTT-3′ for si-THBS4, and 5′-UUCUCCGAACGUGUCACGUTT-3′ and 5′-ACGUGACACGUUCGGAGAATT-3′ for the control siRNA (Invitrogen; Thermo Fisher Scientific, Inc.).

Techniques: Expressing, Transfection, Negative Control