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igfbp6  (R&D Systems)


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    R&D Systems igfbp6
    a) Dot plot representing IGFBP5 (left), <t>IGFBP6</t> (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.
    Igfbp6, supplied by R&D Systems, used in various techniques. Bioz Stars score: 93/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/igfbp+6/Human+IGFBP-6+Antibody/bio_rxiv__64898__2025__12__03__692056-164-38-40
    Average 93 stars, based on 4 article reviews
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    Images

    1) Product Images from "IGFBPs define distinct pro-tumorigenic CAF subtypes in lung cancer tumour-microenvironment"

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

    Journal: bioRxiv

    doi: 10.64898/2025.12.03.692056

    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.
    Figure Legend 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.

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

    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)
    Figure Legend 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)

    Techniques Used: Biomarker Discovery, Knockdown

    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.
    Figure Legend 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.

    Techniques Used: Activity Assay, Knockdown, Western Blot

    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.
    Figure Legend 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.

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

    Related Articles

    Western Blot:

    Article Title: Myelin Formation during Development of the CNS Is Delayed in Matrix Metalloproteinase-9 and -12 Null Mice
    Article Snippet: .. Western blot for IGFBP-6 (goat anti-mouse IGFBP-6; R&D Systems) was then performed. ..

    Membrane:

    Article Title: Matrix metalloproteinase-7 degrades all insulin-like growth factor binding proteins and facilitates insulin-like growth factor bioavailability.
    Article Snippet: Proteolytic modification of insulin-like growth factor binding proteins (IGFBPs) plays an important physiological role in regulating insulin-like growth factor (IGF) bioavailability.. Recently, we demonstrated that matrix metalloproteinase-7 (MMP-7)/Matrilysin produced by various cancer cells catalyzes the proteolysis of IGFBP-3 in vitro and regulates IGF bioavailability, resulting in an anti-apoptotic effect against anchorage-independent culture.. In the present study, we investigated whether MMP-7 contributes to proteolysis of the other five IGFBPs, IGFBP-1, IGFBP-2, IGFBP-4, IGFBP-5, and IGFBP-6, and whether this results in phosphorylation of the IGF type 1 receptor (IGF-1R).

    Article Title: Runx1 promotes angiogenesis by downregulation of insulin-like growth factor-binding protein-3.
    Article Snippet: .. Cells (1.5 105) were resuspended in 1ml of StemPro-34 SFM complete medium (Invitrogen) with or without Dox, recombinant mouse IGFBP3 or IGFBP-6 (R&D Systems) and overlayed on a Biocoat matrigel basement membrane (BD Biosciences) in a six-well plate. ..

    Incubation:

    Article Title: Matrix metalloproteinase-7 degrades all insulin-like growth factor binding proteins and facilitates insulin-like growth factor bioavailability.
    Article Snippet: Proteolytic modification of insulin-like growth factor binding proteins (IGFBPs) plays an important physiological role in regulating insulin-like growth factor (IGF) bioavailability.. Recently, we demonstrated that matrix metalloproteinase-7 (MMP-7)/Matrilysin produced by various cancer cells catalyzes the proteolysis of IGFBP-3 in vitro and regulates IGF bioavailability, resulting in an anti-apoptotic effect against anchorage-independent culture.. In the present study, we investigated whether MMP-7 contributes to proteolysis of the other five IGFBPs, IGFBP-1, IGFBP-2, IGFBP-4, IGFBP-5, and IGFBP-6, and whether this results in phosphorylation of the IGF type 1 receptor (IGF-1R).

    Bioprocessing:

    Article Title: Matrix metalloproteinase-7 degrades all insulin-like growth factor binding proteins and facilitates insulin-like growth factor bioavailability.
    Article Snippet: Proteolytic modification of insulin-like growth factor binding proteins (IGFBPs) plays an important physiological role in regulating insulin-like growth factor (IGF) bioavailability.. Recently, we demonstrated that matrix metalloproteinase-7 (MMP-7)/Matrilysin produced by various cancer cells catalyzes the proteolysis of IGFBP-3 in vitro and regulates IGF bioavailability, resulting in an anti-apoptotic effect against anchorage-independent culture.. In the present study, we investigated whether MMP-7 contributes to proteolysis of the other five IGFBPs, IGFBP-1, IGFBP-2, IGFBP-4, IGFBP-5, and IGFBP-6, and whether this results in phosphorylation of the IGF type 1 receptor (IGF-1R).

    Recombinant:

    Article Title: Runx1 promotes angiogenesis by downregulation of insulin-like growth factor-binding protein-3.
    Article Snippet: .. Cells (1.5 105) were resuspended in 1ml of StemPro-34 SFM complete medium (Invitrogen) with or without Dox, recombinant mouse IGFBP3 or IGFBP-6 (R&D Systems) and overlayed on a Biocoat matrigel basement membrane (BD Biosciences) in a six-well plate. ..

    Enzyme-linked Immunosorbent Assay:

    Article Title: A characteristic signature of insulin-like growth factor (IGF) axis expression during osteogenic differentiation of human dental pulp cells (hDPCs): Potential co-ordinated regulation of IGF action
    Article Snippet: Taqman probes and primers were from ABI (see Supplementary Table 1). .. ELISA kits for assay of IGFBP-2 (#DGB200), IGFBP-3 (#DGB300), IGFBP-4 (#DY804), IGFBP-5 (#DY875), IGFBP-6 (#DY876), IGF1 (#DG100) and IGF2 (#DG200) were from R&D Systems (Abingdon, UK). .. RhIGFBP-4 (#804-GB), rhIGFBP-5 (#875-B5), rhIGF-1(#291-G1) and rhIGF-2 (#292-G2) were also from R&D Systems.



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    Image Search Results


    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.

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    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

    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.

    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

    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.

    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

    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)

    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

    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.

    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

    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.

    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