igfbp6  (Proteintech)

Journal: International Journal of Medical Sciences

Article Title: Stem cells ameliorate neurotrauma-induced visual disturbances and retinal degeneration via broad normalization of β-catenin-related signaling

doi: 10.7150/ijms.123975

Figure Lengend Snippet: MSC therapy enhances growth factor expression and receptor signaling in SI-injured R28 cells and TBI rats. (A) Representative growth factor antibody array images from conditioned media of R28 cells subjected to stretch injury (SI), with or without MSC coculture. Increased secretion of HGF, IGFBP-4, IGFBP-6, and VEGF was observed in the SI+MSC group. Positive controls are marked in red boxes; significantly regulated proteins are highlighted in blue. Quantitative analysis of mean pixel density is shown. (B) ELISA quantification of serum VEGF, HGF, IGFBP-4, and IGFBP-6 levels in TBI rats. TBI reduced systemic levels of these growth factors, while MSC treatment significantly restored them (except IGFBP-6). (C) ELISA quantification of VEGF, HGF, IGFBP-4, and IGFBP-6 in whole eyeball lysates (OD and OS). Due to the technical difficulty in isolating the retina alone, entire ocular globes were homogenized for assay. TBI significantly reduced VEGF, HGF, and IGFBP-6 levels in the eye, while MSC therapy markedly reversed these changes (except VEGF). (D) Western blot analysis of IGF-1R, VEGFR2, phosphorylated VEGFR2 (p-VEGFR2), and c-Met in R28 cell lysates. SI reduced receptor expression, while MSC coculture significantly restored it. Densitometric quantification is normalized to β-actin. (E) Western blot analysis of downstream signaling proteins in R28 cells. MSC coculture upregulated phosphorylated Akt (p-Akt), Sirtuin 1 (SirT1), and β-catenin in SI-injured cells. Quantified values are normalized to β-actin (p < 0.0001). Abbreviations: HGF, hepatocyte growth factor; IGFBP-4/6, insulin-like growth factor binding protein-4/6; VEGF, vascular endothelial growth factor; IGF-1R, insulin-like growth factor 1 receptor; VEGFR2, vascular endothelial growth factor receptor 2; p-Akt, phosphorylated Akt; SirT1, Sirtuin 1. Data are presented as mean ± SD. In vitro experiments (A, D, E) were performed in at least three independent replicates. In vivo data (B, C) are from n=8-9 (serum ELISA) or n = 6 (ocular tissues ELISA) animals per group. For original blot images, see the Supplementary Data file.

Article Snippet: IGFBP-6 , 1:200 , Bioss , BS-4064R , IF.

Techniques: Expressing, Ab Array, Enzyme-linked Immunosorbent Assay, Western Blot, Binding Assay, In Vitro, In Vivo

Journal: International Journal of Medical Sciences

Article Title: Stem cells ameliorate neurotrauma-induced visual disturbances and retinal degeneration via broad normalization of β-catenin-related signaling

doi: 10.7150/ijms.123975

Figure Lengend Snippet: MSC therapy restores TBI-induced neurotrophic growth factor signaling downregulation in retinal neurons. (A) Representative immunofluorescence images of retinal sections (400× magnification) from the indicated groups (Sham+Veh, Sham+MSC, TBI+Veh, TBI+MSC). Images show neuronal co-expression of growth factor ligands and receptors. Top panel: Triple staining for NeuN (neuronal marker, gray), c-Met (HGF receptor, green), and HGF (red). Second panel: NeuN, IGF-1R (IGF receptor, green), and IGFBP-4 (red). Third panel: NeuN, IGF-1R (green), and IGFBP-6 (red). Bottom panel: NeuN, VEGFR2 (VEGF receptor, green), and VEGF (red). DAPI (blue) stains nuclei. Retinal regions include ipsilesional OD-T, OS-N and contralesional OD-N, OS-T segments. (B-E) Quantification of triple-positive cells (NeuN+ growth factor receptor+ ligand) per retinal section: (B) NeuN+c-Met+HGF, (C) NeuN+IGF-1R+IGFBP-4, (D) NeuN+IGF-1R+IGFBP-6, (E) NeuN+VEGFR2+VEGFA. Data were obtained from 6 animals in each group and expressed as mean±SD.

Article Snippet: IGFBP-6 , 1:200 , Bioss , BS-4064R , IF.

Techniques: Immunofluorescence, Expressing, Staining, Marker

Journal: bioRxiv

Article Title: IGFBPs define distinct pro-tumorigenic CAF subtypes in lung cancer tumour-microenvironment

doi: 10.64898/2025.12.03.692056

Figure Lengend Snippet: a) Dot plot representing IGFBP5 (left), IGFBP6 (centre) and IGFBP7 (right) expressions in different cell types in publicly available lung single-cell RNA sequencing datasets. Data was generated from the Curated Cancer Cell Atlas. The size of the circle dot represents the percentage of expressing cells, while the colour represents the mean log2 expression level, as indicated in the scale bar (below), b) mRNA expression (fold change) of IGFBP5, 6 and 7 in patient-derived lung normal fibroblasts (NF) and cancer-associated fibroblasts (CAF). Data are presented as mean ± SEM (n=3 biological triplicates). Statistical analysis was conducted using an unpaired t-test with Welch’s correction; exact p-values are represented in the graph, c) Immunoblots (IBs) showing IGFBP5, 6 and 7 expressions in whole cell lysates of NF and CAF. β-Actin served as the loading control. Densitometric analysis was done using Image Lab (v6.1.0), d) IBs showing secreted and cellular (corresponding serum-starved cell lysates) expression of IGFBP5, 6 and 7. β-Actin served as the loading control for serum-starved cellular lysates, while Ponceau blot served as the loading control for secretory lysates. All IBs are representative of three biological triplicates.

Article Snippet: Cells were lysed directly with RIPA buffer or Laemmli buffer and subjected to SDS PAGE and primary antibodies of α-Smooth Muscle Actin (#19245, CST, 1:1000), FAP (#66562, CST, 1:1000), S100A4 (#13018, CST, 1:1000), IGFBP5 (sc-515184, Santa Cruz, 1:1000), IGFBP6 (AF876, R&D Systems, 1:1000), IGFBP7(sc-365293, Santa Cruz 1:750), Phospho-p44/42 MAPK (Thr202/Tyr204) (#9101, CST, 1:2000), p44/42 MAPK (Thr202/Tyr204) (137F5)(#4695, CST, 1:2000), Phospho-Akt (Ser473) (#4060, CST, 1:2000), Akt (pan) (#2920, CST, 1:2000), Phospho-Insulin R(Y1162/Y1163)/IGF-1R (Y1132/Y1136) (AF2507, R&D Systems, 1:1000), IGF-I R/IGF1R (MAB391, R&D Systems, 1:1000), Phospho-Stat3 (Tyr705) (#9145, CST, 1:2000), Stat3 (#9139, CST, 1:2000), IL6 (#12153, CST, 1:1000), β-Actin (8H10D10) (#3700, CST, 1:2000), GAPDH (#2118, CST, 1:10,000).

Techniques: RNA Sequencing, Generated, Expressing, Derivative Assay, Western Blot, Control

Journal: bioRxiv

Article Title: IGFBPs define distinct pro-tumorigenic CAF subtypes in lung cancer tumour-microenvironment

doi: 10.64898/2025.12.03.692056

Figure Lengend Snippet: a) Volcano plot representing differentially expressed genes between CAF siControl vs siIGFBP5 b) CAF siControl vs siIGFBP6 c) CAF siControl vs siIGFBP7 (n=3 biological replicates). A log2 fold change of 0.5 was set as the threshold. Selected genes for validation are marked in blue. d) Enriched pathways based on the differentially expressed genes upon IGFBP5 knockdown e) IGFBP6 knockdown f) IGFBP7 knockdown g) Heatmap comparing the published CAF subtype gene sets with CAF siControl versus siIGFBP5 h) CAF siControl versus siIGFBP6 i) CAF siControl versus siIGFBP7 (n=2 biological replicates)

Article Snippet: Cells were lysed directly with RIPA buffer or Laemmli buffer and subjected to SDS PAGE and primary antibodies of α-Smooth Muscle Actin (#19245, CST, 1:1000), FAP (#66562, CST, 1:1000), S100A4 (#13018, CST, 1:1000), IGFBP5 (sc-515184, Santa Cruz, 1:1000), IGFBP6 (AF876, R&D Systems, 1:1000), IGFBP7(sc-365293, Santa Cruz 1:750), Phospho-p44/42 MAPK (Thr202/Tyr204) (#9101, CST, 1:2000), p44/42 MAPK (Thr202/Tyr204) (137F5)(#4695, CST, 1:2000), Phospho-Akt (Ser473) (#4060, CST, 1:2000), Akt (pan) (#2920, CST, 1:2000), Phospho-Insulin R(Y1162/Y1163)/IGF-1R (Y1132/Y1136) (AF2507, R&D Systems, 1:1000), IGF-I R/IGF1R (MAB391, R&D Systems, 1:1000), Phospho-Stat3 (Tyr705) (#9145, CST, 1:2000), Stat3 (#9139, CST, 1:2000), IL6 (#12153, CST, 1:1000), β-Actin (8H10D10) (#3700, CST, 1:2000), GAPDH (#2118, CST, 1:10,000).

Techniques: Biomarker Discovery, Knockdown

Journal: bioRxiv

Article Title: IGFBPs define distinct pro-tumorigenic CAF subtypes in lung cancer tumour-microenvironment

doi: 10.64898/2025.12.03.692056

Figure Lengend Snippet: a) Ligand activity analysis showing the list of potential ligands that can regulate differentially expressed genes upon IGFBP5 knockdown in CAF, d) IGFBP6 knockdown, g) IGFBP7 knockdown, b) Ligand-target matrix showing the regulatory potential of potential ligands with CAF siControl vs siIGFBP5, e) CAF siControl vs siIGFBP6, h) CAF siControl vs siIGFBP7, c) Western blot analysis of CAF siControl and siIGFBP5, f) CAF siControl and siIGFBP6, i) CAF siControl and siIGFBP7, after treatment with TGFβ1[10 ng/mL] and IL6 [10 ng/mL] for 48 hours. Densitometric analysis was done using Image Lab (v6.1.0). All IBs are representative of three biological triplicates.

Article Snippet: Cells were lysed directly with RIPA buffer or Laemmli buffer and subjected to SDS PAGE and primary antibodies of α-Smooth Muscle Actin (#19245, CST, 1:1000), FAP (#66562, CST, 1:1000), S100A4 (#13018, CST, 1:1000), IGFBP5 (sc-515184, Santa Cruz, 1:1000), IGFBP6 (AF876, R&D Systems, 1:1000), IGFBP7(sc-365293, Santa Cruz 1:750), Phospho-p44/42 MAPK (Thr202/Tyr204) (#9101, CST, 1:2000), p44/42 MAPK (Thr202/Tyr204) (137F5)(#4695, CST, 1:2000), Phospho-Akt (Ser473) (#4060, CST, 1:2000), Akt (pan) (#2920, CST, 1:2000), Phospho-Insulin R(Y1162/Y1163)/IGF-1R (Y1132/Y1136) (AF2507, R&D Systems, 1:1000), IGF-I R/IGF1R (MAB391, R&D Systems, 1:1000), Phospho-Stat3 (Tyr705) (#9145, CST, 1:2000), Stat3 (#9139, CST, 1:2000), IL6 (#12153, CST, 1:1000), β-Actin (8H10D10) (#3700, CST, 1:2000), GAPDH (#2118, CST, 1:10,000).

Techniques: Activity Assay, Knockdown, Western Blot

Journal: bioRxiv

Article Title: IGFBPs define distinct pro-tumorigenic CAF subtypes in lung cancer tumour-microenvironment

doi: 10.64898/2025.12.03.692056

Figure Lengend Snippet: a) Formalin-fixed paraffin-embedded NSCLC tumour sections were stained with hematoxylin and eosin, Sirius Red, antibodies against α-SMA, IGFBP5, 6 and 7. Cases 1 and 2 represent two different patients. ‘T’ indicates tumour and ‘S’ indicates stroma. The arrow indicates the expression of the respective proteins in fibroblasts; scale bar, 50 μm. b) Representative multiplex immunofluorescence staining images of IGFBP5+, IGFBP6+ and IGFBP7+ CAFs. DAPI (blue), αSMA (green), FAP (pink), PanCK (red), IGFBP5 (grey), IGFBP6 (yellow), IGFBP7 (orange) in human non-small-cell lung cancer tissue sections. Scale bars 50 μm. c) survival between high and low expression of CAF_IGFBP5 gene signatures d) CAF_IGFBP6 gene signatures e) CAF_IGFBP7 gene signatures in TCGA-LUAD datasets, pvalues were obtained from two-sided log-Rank tests.

Article Snippet: Cells were lysed directly with RIPA buffer or Laemmli buffer and subjected to SDS PAGE and primary antibodies of α-Smooth Muscle Actin (#19245, CST, 1:1000), FAP (#66562, CST, 1:1000), S100A4 (#13018, CST, 1:1000), IGFBP5 (sc-515184, Santa Cruz, 1:1000), IGFBP6 (AF876, R&D Systems, 1:1000), IGFBP7(sc-365293, Santa Cruz 1:750), Phospho-p44/42 MAPK (Thr202/Tyr204) (#9101, CST, 1:2000), p44/42 MAPK (Thr202/Tyr204) (137F5)(#4695, CST, 1:2000), Phospho-Akt (Ser473) (#4060, CST, 1:2000), Akt (pan) (#2920, CST, 1:2000), Phospho-Insulin R(Y1162/Y1163)/IGF-1R (Y1132/Y1136) (AF2507, R&D Systems, 1:1000), IGF-I R/IGF1R (MAB391, R&D Systems, 1:1000), Phospho-Stat3 (Tyr705) (#9145, CST, 1:2000), Stat3 (#9139, CST, 1:2000), IL6 (#12153, CST, 1:1000), β-Actin (8H10D10) (#3700, CST, 1:2000), GAPDH (#2118, CST, 1:10,000).

Techniques: Formalin-fixed Paraffin-Embedded, Staining, Expressing, Multiplex Assay, Immunofluorescence