Journal: Oncotarget

Article Title: IGFBP6 is a novel nasopharyngeal carcinoma prognostic biomarker

doi: 10.18632/oncotarget.11886

Figure Lengend Snippet: IGFBP6 was detected by immunostaining in primary NPC tissues (magnification ×200). Left:, IGFBP6 positive staining; Middle: IGFBP6 negative staining; Right: isotype control staining ( A ) IGFBP6 mRNA was measured in five NPC cell lines (CNE2, CNE1, SUNE1, HK1 and HONE1) via RT-PCR, with GAPDH as an internal control ( B ) Data are representative of three separate experiments. Western blotting of whole-cell lysates to detect IGFBP6 ( C ) IGFBP6 levels in CM from NPC cells as measured by ELISA ( D ) Data are representative of two separate experiments. All data represent means ± SD from triplicates.

Article Snippet: Recombinant human IGFBP6 (rhIGFBP6) and the anti-human IGFBP6 monoclonal antibody were purchased from R&D Systems (Minneapolis, MN).

Techniques: Immunostaining, Staining, Negative Staining, Control, Reverse Transcription Polymerase Chain Reaction, Western Blot, Enzyme-linked Immunosorbent Assay

Journal: Oncotarget

Article Title: IGFBP6 is a novel nasopharyngeal carcinoma prognostic biomarker

doi: 10.18632/oncotarget.11886

Figure Lengend Snippet: Correlation of IGFBP6 expression with clinical characteristics in patients with NPC

Article Snippet: Recombinant human IGFBP6 (rhIGFBP6) and the anti-human IGFBP6 monoclonal antibody were purchased from R&D Systems (Minneapolis, MN).

Techniques: Expressing

Journal: Oncotarget

Article Title: IGFBP6 is a novel nasopharyngeal carcinoma prognostic biomarker

doi: 10.18632/oncotarget.11886

Figure Lengend Snippet: Multivariate Cox regression analysis for survival prognostic factors in advanced nasopharyngeal carcinoma

Article Snippet: Recombinant human IGFBP6 (rhIGFBP6) and the anti-human IGFBP6 monoclonal antibody were purchased from R&D Systems (Minneapolis, MN).

Techniques: Expressing

Journal: Oncotarget

Article Title: IGFBP6 is a novel nasopharyngeal carcinoma prognostic biomarker

doi: 10.18632/oncotarget.11886

Figure Lengend Snippet: CNE2 (upper panel) and HK1 (lower panel) cell proliferation as measured by MTS assay ( A ) Data represent means ± SD from six wells. * P < 0.05 compared to controls (IGFBP6 0 ng/ml). In transwell assays (upper panel), exogenous IGFBP6 inhibited CNE2 and HK1 cell invasion compared to controls ( B ) Invasive Index (%) was calculated (lower panel) according to the manufacturer's instructions. Columns, means of triplicate assays; bars, SE. ** P < 0.01 compared to controls.

Article Snippet: Recombinant human IGFBP6 (rhIGFBP6) and the anti-human IGFBP6 monoclonal antibody were purchased from R&D Systems (Minneapolis, MN).

Techniques: MTS Assay

Journal: Oncotarget

Article Title: IGFBP6 is a novel nasopharyngeal carcinoma prognostic biomarker

doi: 10.18632/oncotarget.11886

Figure Lengend Snippet: CNE2 cells were stably transfected with IGFBP6-shRNA. Real-time RT-PCR confirmed knockdown efficiency ( A ) Bars, ± SE. Data are representative of three separate experiments. Western blotting confirmed IGFBP6 knockdown ( B ) IGFBP6 knockdown induced tumor cell proliferation compared to controls ( C ) Representative wound-healing assay images ( D ) IGFBP6 knockdown increased tumor cell migration ( E ) Data represent means ± SD. * P < 0.05 compared to controls. Western blotting revealed GSK3β/β-catenin/cylin D1 pathway activation as a result of IGFBP6 knockdown ( F ).

Article Snippet: Recombinant human IGFBP6 (rhIGFBP6) and the anti-human IGFBP6 monoclonal antibody were purchased from R&D Systems (Minneapolis, MN).

Techniques: Stable Transfection, Transfection, shRNA, Quantitative RT-PCR, Knockdown, Western Blot, Wound Healing Assay, Migration, Activation Assay

Journal: Oncotarget

Article Title: IGFBP6 is a novel nasopharyngeal carcinoma prognostic biomarker

doi: 10.18632/oncotarget.11886

Figure Lengend Snippet: Silencing IGFBP6 expression in CNE2 cells promotes tumor metastasis in a mouse model

Article Snippet: Recombinant human IGFBP6 (rhIGFBP6) and the anti-human IGFBP6 monoclonal antibody were purchased from R&D Systems (Minneapolis, MN).

Techniques: Expressing

Journal: Communications Biology

Article Title: Pantothenic acid ameliorates hepatic fibrosis by targeting IGFBP6 to regulate the TGF-β/SMADs pathway

doi: 10.1038/s42003-025-08527-5

Figure Lengend Snippet: a Volcano plot showing the distribution of DEGs. Yellow dots indicate significantly up-regulated genes, green dots indicate significantly down-regulated genes, and pink dots indicate non-differentially expressed genes. b Heat map of relevant DEGs between fibrosis and control groups. c Top 20 most significantly differentially expressed genes, with IGFBP6 showing the highest significance. d Validation using dataset GSE130123 . CCL4-induced 4 weeks hepatic fibrosis model in mice. (control group n = 4, and fibrosis group n = 5). e IHC staining of COL1A1 and IGFBP6 in hepatic sections from control mice and mice at 4 or 8 weeks after intraperitoneal TAA injection (scale bar: 25 μm). Schematic diagram generated by Biorender (agreement number: AR28HM9B0Q). f Primary HEPs ( n = 6) and HSCs ( n = 6) were isolated from the same liver. The mRNA expression of mouse Igfbp6 . g Primary HSCs were isolated from WT mice treated with three doses of vehicle (PBS, n = 6) or TAA (100 mg/kg, n = 6) every 3 days. Igfbp6 mRNA was measured. h scRNA-seq dataset ( GSE221481 ) from mice with uninjured liver or TAA-induced hepatic fibrosis was analyzed for the expression of Igfbp6 . The fraction of cells expressing Igfbp6 in different cell lineages in liver. Data are expressed as mean ± SD, unpaired Student’s t test, two-tailed.

Article Snippet: Recombinant human IGFBP6 protein with His-tag (MCE, HY- P73141 ) was purified using Ni-NTA affinity chromatography (Thermo Scientific, HisPur).

Techniques: Control, Biomarker Discovery, Immunohistochemistry, Injection, Generated, Isolation, Expressing, Two Tailed Test

Journal: Communications Biology

Article Title: Pantothenic acid ameliorates hepatic fibrosis by targeting IGFBP6 to regulate the TGF-β/SMADs pathway

doi: 10.1038/s42003-025-08527-5

Figure Lengend Snippet: a qRT-PCR analysis of Acta2 , Fn1 , Col1a1 , and Igfbp6 mRNA levels and b Western blotting analysis of FN, COL1A1, α-SMA, and IGFBP6 protein expression in hepatic tissue homogenates from control and TAA-treated mice at 4 and 8 weeks ( n = 6), Data are expressed as means ± SD, One-way ANOVA. c qRT-PCR analysis of Acta2 , Fn1 , Col1a1 and Igfbp6 mRNA levels in primary mouse HSCs stimulated with TGF-β (10 ng/mL) for 12 h and 24 h or unstimulated ( n = 3), Data are expressed as means ± SD, One-way ANOVA. d Western blotting analysis of protein expression levels of FN, COL1A1, α-SMA and IGFBP6 in primary mouse HSCs stimulated with TGF-β for 24 and 48 h or unstimulated. e –g Western blotting confirmed IGFBP6-knockout in LX2 cells line ( e ). Fibrosis-related gene and protein expression in LX2 cells line stimulated with TGF-β (10 ng/mL) with or without IGFBP6-knockout detected by ( f ) qRT-PCR and ( g ) Western blotting ( n = 3). Data are expressed as means ± SD, Tow-way ANOVA.

Article Snippet: Recombinant human IGFBP6 protein with His-tag (MCE, HY- P73141 ) was purified using Ni-NTA affinity chromatography (Thermo Scientific, HisPur).

Techniques: Quantitative RT-PCR, Western Blot, Expressing, Control, Knock-Out

Journal: Communications Biology

Article Title: Pantothenic acid ameliorates hepatic fibrosis by targeting IGFBP6 to regulate the TGF-β/SMADs pathway

doi: 10.1038/s42003-025-08527-5

Figure Lengend Snippet: a SwissADME analysis of 30 FDA-approved compounds related to IGFBP6 showing: Heatmap of five drug-likeness rules (Lipinski, Ghose, Veber, Egan, Muegge) and eight pharmacokinetic parameters (GI absorption, BBB permeation, P-gp substrate, CYP1A2/2C19/2C9/2D6/3A4 inhibition). b Bioavailability scores of the compounds. Effect of five candidate compounds on fibrosis-related indicators in TGF-β (10 ng/mL) stimulated primary mouse HSCs were detected ( c ) at 24 h for mRNA levels and ( d ) at 48 h for protein levels ( n = 3). e Chemical structural of PA. f The effect of PA on cell viability of LX2 cells viability was measured using the CCK8 assay ( n = 5). The effect of concentration gradients (0 μM, 0.1 μM, 1 μM, and10μM) of PA against FN, COL1A1, ACTA2, and IGFBP6 in TGF-β (10 ng/mL) -stimulated LX2 cells was examined using ( g ) qRT-PCR for mRNA levels and ( h ) Western blotting for protein levels ( n = 3). Data are expressed as means ± SD, Two-way ANOVA.

Article Snippet: Recombinant human IGFBP6 protein with His-tag (MCE, HY- P73141 ) was purified using Ni-NTA affinity chromatography (Thermo Scientific, HisPur).

Techniques: Inhibition, CCK-8 Assay, Concentration Assay, Quantitative RT-PCR, Western Blot

Journal: Communications Biology

Article Title: Pantothenic acid ameliorates hepatic fibrosis by targeting IGFBP6 to regulate the TGF-β/SMADs pathway

doi: 10.1038/s42003-025-08527-5

Figure Lengend Snippet: a Three-dimensional binding structure of PA (green) with IGFBP6 (gray ribbon). The Pi-H interactions are depicted as magenta dashed lines. b MST analysis of His-tagged IGFBP6 binding to PA ( n = 3). c CETSA melting curves and derived thermal stability profiles ( n = 3). d Western blotting detection of protein expression levels of FN, COL1A1 and α-SMA in IGFBP6-knockout LX2 cells by PA (10 μM) intervention. The effect of PA’s inhibitory action on TGF-β-induced IGFBP6 overexpression in LX2 cells was assessed by Western blotting in the presence or absence of ( e ) the proteasome inhibitor MG132 or ( f ) the autophagy inhibitor 3-MA ( n = 3). Data are expressed as means ± SD, Two-way ANOVA. g LX2 cells were treated with 1 µM PA in the presence or absence of MG132 for 12 h. IP detection was performed with anti-IGFBP6 antibody, and expression of ubiquitin-coupled IGFBP6 was detected with anti-UB antibody ( n = 3). Data are expressed as means ± SD, Two-way ANOVA.

Article Snippet: Recombinant human IGFBP6 protein with His-tag (MCE, HY- P73141 ) was purified using Ni-NTA affinity chromatography (Thermo Scientific, HisPur).

Techniques: Binding Assay, Derivative Assay, Western Blot, Expressing, Knock-Out, Over Expression, Ubiquitin Proteomics

Journal: Communications Biology

Article Title: Pantothenic acid ameliorates hepatic fibrosis by targeting IGFBP6 to regulate the TGF-β/SMADs pathway

doi: 10.1038/s42003-025-08527-5

Figure Lengend Snippet: a Enrichment analysis results after collection of PA targets via Pharmmapper database and GeneCards database. b Western blotting assay for the influence of PA (10 μM) on different times of TGF-β (10 ng/mL) stimulation-initiated SMAD2/3 phosphorylation levels in primary mouse HSCs ( n = 3). c Effects of different concentrations of PA (0 μM, 0.1 μM, 1 μM, and10 μM) on the phosphorylation level of SMAD2/3 induced by TGF-β (10 ng/mL) action in primary HSCs of mice analyzed by Western blotting ( n = 3). Effect of PA on phosphorylation levels of SMAD2/3 and IGFBP6 in TAA-induced fibrotic hepatic mice in vivo by ( d ) Western blotting and ( e ) IHC ( n = 6, scale bar: 50 μm).

Article Snippet: Recombinant human IGFBP6 protein with His-tag (MCE, HY- P73141 ) was purified using Ni-NTA affinity chromatography (Thermo Scientific, HisPur).

Techniques: Western Blot, Phospho-proteomics, In Vivo

Journal: Communications Biology

Article Title: Pantothenic acid ameliorates hepatic fibrosis by targeting IGFBP6 to regulate the TGF-β/SMADs pathway

doi: 10.1038/s42003-025-08527-5

Figure Lengend Snippet: Phospho-SMAD2/3 levels detected by Western blotting in ( a ) control group and ( b ) IGFBP6-knockout group treated with TGF-β (10 ng/mL) for 15 min, 30 min, or 60 min with or without PA (10 µM) ( n = 3). Data are expressed as means ± SD, One-way ANOVA. c Immunofluorescence analysis of TGF-β (10 ng/mL) -induced SMAD2/3 nuclear translocation in LX2 cells with or without IGFBP6 knockout, treated with or without PA (10 µM) (scale bar: 25 μm). d The mRNA expression levels of Acta2 , Fn1 , and Col1a1 in primary mouse HSCs in the presence or absence of PA or the SMAD3 inhibitor SIS3 were detected by qRT-PCR ( n = 3). Data are expressed as means ± SD, Two-way ANOVA.

Article Snippet: Recombinant human IGFBP6 protein with His-tag (MCE, HY- P73141 ) was purified using Ni-NTA affinity chromatography (Thermo Scientific, HisPur).

Techniques: Western Blot, Control, Knock-Out, Immunofluorescence, Translocation Assay, Expressing, Quantitative RT-PCR

Journal: Molecular Pain

Article Title: Global transcriptome analysis of rat dorsal root ganglia to identify molecular pathways involved in incisional pain

doi: 10.1177/1744806920956480

Figure Lengend Snippet: List of upregulated genes in the dorsal root ganglia after plantar incision.

Article Snippet: Taqman gene expression assays include Rn00565371_m1 (IGFBP6), Rn01454518_m1 (IGF2), and Rn00667869_m1 (Actb).

Techniques: Clinical Proteomics, Membrane, Binding Assay, RNA Binding Assay

Journal: Molecular Pain

Article Title: Global transcriptome analysis of rat dorsal root ganglia to identify molecular pathways involved in incisional pain

doi: 10.1177/1744806920956480

Figure Lengend Snippet: Altered IGF1 signaling in DRG of rats after plantar incision. (a, b) IGF1 gene networks showing altered expression level in incision group (a) and incision with capsaicin pretreated group (b). Boxed genes indicate those that responded to capsaicin treatment. (c) IPA mapped differentially expressed genes onto IGF1 canonical pathway suggesting reduced cell growth and proliferation. (d) qPCR validation of IGF2 and IGFBP6 in DRG. Values are mean ± SD, n = 3–6/group, t-test, **p < 0.01, ***p < 0.001.

Article Snippet: Taqman gene expression assays include Rn00565371_m1 (IGFBP6), Rn01454518_m1 (IGF2), and Rn00667869_m1 (Actb).

Techniques: Expressing, Biomarker Discovery

Journal: Neurological research

Article Title: The clinical characteristics and prognostic value of IGFBP6 in glioma.

doi: 10.1080/01616412.2021.1963620

Figure Lengend Snippet: Figure 1. The expression of IGFBP6 according to WHO grades, IDH status and 1p/19q status in CGGA and TCGA datasets. (a-c) IGFBP6 was significantly increased in WHO IV, IDH wild type and 1p/19q intact glioma samples in TCGA dataset (d-f). IGFBP6 was significantly increased in WHO IV, IDH wild type and 1p/19q intact glioma samples in CGGA dataset. Photographs of immuno histochemical staining of IGFBP6 in different grades of gliomas. Positive cells are stained brown. (g) Diffuse astrocytoma (WHO grade II). (h) Anaplastic astrocytoma (WHO grade III). (i) Glioblastoma multiforme (WHO grade IV). Magnification, x200. *, **, *** and **** indicate P < 0.05, P < 0.01, P < 0.001 and P < 0.0001, respectively.

Article Snippet: IGFBP6 Monoclonal Antibody (67,567-1-Ig, dilution: 1:200, Proteintech) was used to detect IGFBP6 protein expression.

Techniques: Expressing, Staining

Journal: Neurological research

Article Title: The clinical characteristics and prognostic value of IGFBP6 in glioma.

doi: 10.1080/01616412.2021.1963620

Figure Lengend Snippet: Figure 2. The difference of IGFBP6 in four TCGA molecular markers. (a, c) The expression of IGFBP6 was significantly upregulated in mesenchymal subtype in TCGA and CGGA datasets. (b, d) The ROC curves indicated that IGFBP6 expression exhibited highly sensitivity and specificity to predict mesenchymal subtype in TCGA and CGGA datasets. Area under curve (AUC) was 0.831 and 0.858, respectively. (e) Photographs of immunohistochemical staining of IGFBP6 in gliomas with different molecular subtypes. Positive cells are stained brown. Magnification, x200.

Article Snippet: IGFBP6 Monoclonal Antibody (67,567-1-Ig, dilution: 1:200, Proteintech) was used to detect IGFBP6 protein expression.

Techniques: Expressing, Immunohistochemical staining, Staining

Journal: Neurological research

Article Title: The clinical characteristics and prognostic value of IGFBP6 in glioma.

doi: 10.1080/01616412.2021.1963620

Figure Lengend Snippet: Figure 3. The gene ontology analysis of IGFBP6. (a, b) Heatmaps show the IGFBP6-related genes in CGGA and TCGA datasets. (c) The IGFBP6 expression was positively related with immune responses, immune cell migration, regulation of JAK-STAT pathway and so on. (d) The IGFBP6 expression was negatively related with the normal physiological functions of neurons and cells.

Article Snippet: IGFBP6 Monoclonal Antibody (67,567-1-Ig, dilution: 1:200, Proteintech) was used to detect IGFBP6 protein expression.

Techniques: Expressing, Migration

Journal: Neurological research

Article Title: The clinical characteristics and prognostic value of IGFBP6 in glioma.

doi: 10.1080/01616412.2021.1963620

Figure Lengend Snippet: Figure 4. IGFBP6 related checkpoints and inflammation activities in CGGA and TCGA cohorts. (a, b) IGFBP6 expression was positively correlated with PDCD1, PDCD1L2, HAVCR2 and CD274 expression. (c, d) The relationship between IGFBP6 expression and seven inflammation activities. In pie charts, positive correlations are displayed in green and negative correlations in red. Color intensity and the size of the circle are proportional to the correlation coefficients.

Article Snippet: IGFBP6 Monoclonal Antibody (67,567-1-Ig, dilution: 1:200, Proteintech) was used to detect IGFBP6 protein expression.

Techniques: Expressing

Journal: Neurological research

Article Title: The clinical characteristics and prognostic value of IGFBP6 in glioma.

doi: 10.1080/01616412.2021.1963620

Figure Lengend Snippet: Figure 5. IGFBP6 was a prognostic factor in glioma patients. (a-f) Kaplan–Meier survival analysis showed that high expression of IGFBP6 indicated a significantly worse OS in all grade glioma patients (p < 0.0001), GBM patients (p = 0.029), IDH mutant patients (p = 0.01) and IDH wild type (p = 0.015) patients in CGGA dataset. The lower level of IGFBP6 significantly increased glioma sensitivity to radiotherapy (p = 0.002) or chemotherapy (p = 0.034) in CGGA dataset. (g-j) Kaplan–Meier survival analysis showed that high expression of IGFBP6 indicated a significantly worse OS in all grade glioma patients (p < 0.0001), GBM patients (p = 0.0061), IDH mutant patients (p = 0.0283) and IDH wild type (p = 0.0037) patients in TCGA dataset.

Article Snippet: IGFBP6 Monoclonal Antibody (67,567-1-Ig, dilution: 1:200, Proteintech) was used to detect IGFBP6 protein expression.

Techniques: Expressing, Mutagenesis

Journal: Journal of the American Heart Association: Cardiovascular and Cerebrovascular Disease

Article Title: Endothelial Dysfunction Contributes to Chondrocyte Senescence Associated With Insulin‐Like Growth Factor‐Binding Protein‐6 in a Spontaneously Hypertensive Rat Model

doi: 10.1161/JAHA.124.042132

Figure Lengend Snippet: A , Schematic for isolation of primary endothelial cells for RNA‐sequencing. B , Volcano plot of differentially expressed genes between WKY and SHR endothelial cells. Genes are considered significantly different when adjusted P value is <0.05 and fold change is >1. Red: higher expression in SHR. Blue: higher expression in WKY. Green: not significantly different. Top upregulated and downregulated genes were annotated. Note that FMO1 is among the annotated upregulated genes. C , Top upregulated Gene Ontology terms pertaining to molecular function in SHR compared with WKY. (n=3). D and E , Heatmap of genes involved in monooxygenase activity and the normalized expression of selected genes, FMO1, FMO2, and FMO3. F through G , Heatmap of genes in insulin‐like growth factor system and the normalized expression of selected genes, IGFBP6. Gene expression was normalized using R package DESeq2’s median of ratios. Adjusted P value was shown for selected genes of interest. Likelihood‐ratio test included in DESeq2 was used to compute the adjusted P value. All data were presented as mean±SE. * P <0.05; ** P <0.01; *** P <0.001; **** P <0.0001. EC indicates endothelial cells; FMO, flavin‐containing monooxygenase; IGFBP6, insulin‐like growth factor‐binding protein 6; ns, not significant; SHR, spontaneously hypertensive rats; and WKY, Wistar Kyoto rats.

Article Snippet: Human IGFBP6 protein (1 μg/mL, HY‐ P73141 , MCE) was administered to C28/I2 cell culture for 6 hours to induce cellular senescence.

Techniques: Isolation, RNA Sequencing, Expressing, Activity Assay, Gene Expression, Binding Assay

Journal: Journal of the American Heart Association: Cardiovascular and Cerebrovascular Disease

Article Title: Endothelial Dysfunction Contributes to Chondrocyte Senescence Associated With Insulin‐Like Growth Factor‐Binding Protein‐6 in a Spontaneously Hypertensive Rat Model

doi: 10.1161/JAHA.124.042132

Figure Lengend Snippet: A and B , Representative immunofluorescent staining and quantification of 4HNE in aortas in 9‐month‐old rats. (N=6, male). C and D , Representative images and quantification of p16ink4a staining in aorta in 9‐month‐old animals. Scale bar 50 μm. (N=6, male). E , Carotid artery stiffness in 9‐month‐old WKY and SHR. F , Representative immunofluorescent staining of 4HNE and CD31 in synovial tissue in 9‐month‐old WKY and SHR. (N=6, male). G , Photoacoustic images and measurement of oxygen saturation in joint tissue in 9‐month‐old WKY and SHR. (N=6, male). H and I , Immunohistochemistry staining and quantification of IGFBP6 in the synovium of 9‐month‐old WKY and SHR. (N=6, male). J and M , Representative images and quantification of α‐SMA and Masson’s Trichrome (blue) staining in synovium of 9‐month‐old WKY and SHR. (N=6, male). N and O , Immunohistochemistry staining and quantification of p16ink4a in synovium of 9‐month‐old WKY and SHR. (N=6, male). P and Q , Immunohistochemistry staining and quantification of IGFBP6 in subchondral bone of 9‐month‐old WKY and SHR. (N=6, male). R and S , Representative images and quantification of micro‐computed tomography result for subchondral bone of WKY and SHR joint for 9‐month‐old WKY and SHR. (N=6, male) T , IGFBP6 staining of cartilage in 9‐month‐old WKY and SHR. U , Representative images of Safranin‐O/Fast Green staining for 9‐month‐old WKY and SHR joints. (N=6, male) V , OARSI score grading system was adopted from Waldstein et al. ²⁰ (N=6, male) W and X , Immunohistochemistry staining and quantification of p16ink4a in cartilage of 9‐month‐old WKY and SHR. (N=6, male). Y and Z , Immunohistochemistry staining and quantification of MMP13 in cartilage of 9‐month‐old WKY and SHR. (N=6, male) All data were presented as mean±SE. Two‐tailed Student’s t test (unpaired) was performed between WKY and SHR. * P <0.05; ** P <0.005; *** P <0.001. α‐SMA indicates alpha smooth muscle actin; BMD, bone mineral density; IGFBP6, insulin‐like growth factor‐binding protein 6; MMP13, matrix metalloproteinase 13; OARSI, Osteoarthritis Research Society International; SHR, spontaneously hypertensive rats; and WKY, Wistar Kyoto rats.

Article Snippet: Human IGFBP6 protein (1 μg/mL, HY‐ P73141 , MCE) was administered to C28/I2 cell culture for 6 hours to induce cellular senescence.

Techniques: Staining, Immunohistochemistry, Micro-CT, Two Tailed Test, Binding Assay

Journal: Journal of the American Heart Association: Cardiovascular and Cerebrovascular Disease

Article Title: Endothelial Dysfunction Contributes to Chondrocyte Senescence Associated With Insulin‐Like Growth Factor‐Binding Protein‐6 in a Spontaneously Hypertensive Rat Model

doi: 10.1161/JAHA.124.042132

Figure Lengend Snippet: A , Divided the 17 proteins of interest into 2 groups to analyze their functional enrichment according to their response to H 2 O 2 challenge. B , Heatmap showing relative expression of proteins across groups. (n=4). C and D , SA‐β‐gal staining in C28/I2 cells with or without IGFBP6. (n=3). E , Relative mRNA level of p16 INK4a and p21 in C28/I2 cells with or without IGFBP6. (n=3) ( F and G ) SA‐β‐gal staining in C28/I2 cell for control, H 2 O 2 , IGFBP6 antibody, and H 2 O 2 +IGFBP6 antibody group. (n=3). H , Relative mRNA level of p16 INK4a for control, H 2 O 2 , IGFBP6 antibody, and H 2 O 2 + IGFBP6 antibody group. (n=3) All data are shown as means±SE, and each data point represents an individual experiment. One‐way ANOVA with Tukey’s multiple‐comparisons test was employed for statistical analysis to compare between each experimental group and control group. * P <0.05; ** P <0.005; *** P <0.001. ECM indicates extracellular matrix; IGFBP6, insulin‐like growth factor‐binding protein 6; and SA‐β‐gal, senescence‐associated beta‐galactosidase.

Article Snippet: Human IGFBP6 protein (1 μg/mL, HY‐ P73141 , MCE) was administered to C28/I2 cell culture for 6 hours to induce cellular senescence.

Techniques: Functional Assay, Expressing, Staining, Control, Binding Assay

Journal: Journal of the American Heart Association: Cardiovascular and Cerebrovascular Disease

Article Title: Endothelial Dysfunction Contributes to Chondrocyte Senescence Associated With Insulin‐Like Growth Factor‐Binding Protein‐6 in a Spontaneously Hypertensive Rat Model

doi: 10.1161/JAHA.124.042132

Figure Lengend Snippet: A , Flow diagrams on grouping and experimental design for evaluation of the therapeutic effects of captopril treatment. B , Systolic blood pressure in SHR and SHR after captopril treatment. C and D , Immunohistochemistry staining and quantification of p16ink4a in aorta of captopril‐treated and saline‐treated 9‐month‐old SHR. (N=6, 3 male and 3 female). E , Representative immunofluorescent staining of 4HNE and CD31 in synovial vasculature in 9‐month‐old SHR and SHR after captopril treatment. F and G , Immunohistochemistry staining and quantification of IGFBP6 in synovium of captopril‐treated and saline‐treated 9‐month‐old SHR. (N=6, 3 male and 3 female). H and K , Representative images and quantification of α‐SMA and Masson’s Trichrome staining in synovium of 9‐month‐old SHR and SHR after captopril treatment. (N=6, 3 male and 3 female). L and M , Immunohistochemistry staining and quantification of p16ink4a in synovium of captopril‐treated and saline‐treated 9‐month‐old SHR. (N=6, 3 male and 3 female). N through O , Immunohistochemistry staining and quantification of IGFBP6 in bone of captopril‐treated and saline‐treated 9‐month‐old SHR and SHR after captopril treatment. (N=6, 3 male and 3 female). P and Q , Representative images and quantification of micro‐computed tomography result for subchondral bone of joint for 9‐month‐old SHR and SHR after captopril treatment. (N=6, N=6, 3 male and 3 female). R , Representative images of Safranin‐O/Fast Green staining for 9‐month‐old joints of SHR and SHR after captopril treatment. (N=6, 3 male and 3 female). S , OARSI score for treated and saline‐treated 9‐month‐old SHR and SHR after captopril treatment. (N=6, 3 male and 3 female). T and U , Immunohistochemistry staining and quantification of p16ink4a in cartilage of captopril‐treated and saline‐treated 9‐month‐old SHR. (N=6, 3 male and 3 female). V and W , Immunohistochemistry staining and quantification of MMP13 in cartilage of captopril‐treated and saline‐treated 9‐month‐old SHR. Scale bar 50 μm. (N=6, 3 male and 3 female) All data were presented as mean±SE. Two‐tailed Student’s t test (unpaired) was performed for staining intensity and percentage of positively stained cells between SHR and captopril‐treated group. * P <0.05; ** P <0.005; *** P <0.001. α‐SMA indicates alpha smooth muscle actin; BMD, bone mineral density; IGFBP6, insulin‐like growth factor‐binding protein 6; MMP13, matrix metalloproteinase 13; OARSI, Osteoarthritis Research Society International; and SHR, spontaneously hypertensive rats.

Article Snippet: Human IGFBP6 protein (1 μg/mL, HY‐ P73141 , MCE) was administered to C28/I2 cell culture for 6 hours to induce cellular senescence.

Techniques: Immunohistochemistry, Staining, Saline, Micro-CT, Two Tailed Test, Binding Assay

Journal: Cancer science

Article Title: Matrix metalloproteinase-7 triggers the matricrine action of insulin-like growth factor-II via proteinase activity on insulin-like growth factor binding protein 2 in the extracellular matrix.

doi: 10.1111/j.1349-7006.2007.00448.x

Figure Lengend Snippet: Fig. 1. Expression of insulin-like growth factor (IGF)-II and insulin-like growth factor binding proteins (IGFBPs) in HT29 cells. Ten micrograms of trichloroacetic acid-precipitable protein from conditioned medium of HT29 cells were fractionated on a 15% sodium dodecylsulfate–polyacrylamide gel electrophoresis (SDS-PAGE) and analyzed by Western blotting for the expression levels of IGF-II and all IGFBPs (IGFBP-1 to IGFBP-6). Arrow of each left lane indicates positive control (100 ng of recombinant protein, respectively).

Article Snippet: Recombinant human IGF-II and recombinant human IGFBPs (IGFBP-1 to IGFBP-6) were obtained from R & D Systems (Minneapolis, MN, USA).

Techniques: Expressing, Binding Assay, Polyacrylamide Gel Electrophoresis, SDS Page, Western Blot, Positive Control, Recombinant