Journal: bioRxiv

Article Title: A Wnt-responsive fibrocartilage progenitor system coordinates postnatal mandibular condylar cartilage growth

doi: 10.64898/2026.03.25.714159

Figure Lengend Snippet: (A) Differential gene expression analysis was performed comparing H2B–EGFP –positive and H2B–EGFP –negative cells within the MC-progenitor cluster identified by single-cell RNA sequencing. Genes are ranked by statistical significance. Foxm1 is significantly enriched in the H2B–EGFP –positive population, whereas Tgfb1 is enriched in the H2B–EGFP –negative population, indicating divergent transcriptional programs associated with Wnt activity. (B) Feature plots were generated to visualize expression of representative genes across mandibular condylar cartilage populations. Wnt-responsive cells show enriched expression of Foxm1 and IGF signaling–related genes ( Igf1 , Igf1r , Igfbp4 , Igfbp7 ), whereas Wnt-low populations express Tgfb1 and related factors ( Igf2r , Igfbp5 , Igfbp6 ), supporting distinct signaling states. (C) Western blot analysis was performed in isolated Wnt-responsive cells transfected with control vector or constitutively active β-catenin (S33Y). Cells were stimulated with recombinant IGF1 for the indicated time points. β-catenin activation enhances Foxm1 expression and downstream mitogenic signaling, including ERK and IGF1R phosphorylation, indicating that β-catenin promotes proliferative signaling responses. (D) Co-immunoprecipitation was performed to assess interaction between β-catenin and Foxm1. Cell lysates immunoprecipitated with anti–β-catenin antibody show enrichment of Foxm1 compared with control IgG, indicating a physical association between β-catenin and Foxm1. (E,F) Histological and immunofluorescence analyses were performed on mandibular condyles from control and Axin2 CreERT2 ;Ctnnb1 fl/+ ;Foxm1 fl/+ compound heterozygous mice at P42. H&E staining reveals reduced fibrocartilage thickness, and Ki67 staining shows decreased proliferative activity, indicating cooperative effects of β-catenin and Foxm1 in maintaining fibrocartilage proliferation. Scale bar, 100 μm. (G) Quantification of fibrocartilage thickness and Ki67-positive cells was performed. Compound heterozygous mice show reduced fibrocartilage thickness and decreased proliferation compared with controls. Data are presented as mean ± s.d. Each dot represents one biologically independent animal. (H,I) Histological and immunofluorescence analyses were performed on mandibular condyles from control and Axin2 CreERT2 ;Foxm1 fl/fl mice at P42. Foxm1 deletion results in marked condylar hypoplasia and reduced proliferative activity, indicating a critical role for Foxm1 in fibrocartilage growth. Scale bar, 100 μm. (J) Quantification of cartilage thickness and proliferative indices was performed in Foxm1 conditional knockout mice. Foxm1 deficiency significantly reduces cartilage growth and proliferation. Data are presented as mean ± s.d. Each dot represents one biologically independent animal. Statistical significance was assessed using two-tailed Student’s t-test. n.s., not significant; **P < 0.01; ****P < 0.0001. Abbreviations: sz, superficial zone; fc, fibrocartilage zone; cc, chondrocartilage zone.

Article Snippet: Cells were stimulated with recombinant mouse IGF1 (Cat No. 791-MG-050, R&D systems) for 0, 30, 60, or 180 min.

Techniques: Gene Expression, Single Cell, RNA Sequencing, Activity Assay, Generated, Expressing, Western Blot, Isolation, Transfection, Control, Plasmid Preparation, Recombinant, Activation Assay, Phospho-proteomics, Immunoprecipitation, Immunofluorescence, Staining, Knock-Out, Two Tailed Test

Journal: bioRxiv

Article Title: A Wnt-responsive fibrocartilage progenitor system coordinates postnatal mandibular condylar cartilage growth

doi: 10.64898/2026.03.25.714159

Figure Lengend Snippet: (A) RNAscope in situ hybridization showing Foxm1 transcript localization within the fibrocartilage compartment of the mandibular condyle. (B) RNAscope detection of Igf1 transcripts enriched in the superficial region of the fibrocartilage layer. (C) Violin plot comparing Foxm1 expression between H2B-EGFP –positive and H2B-EGFP –negative cells within the MC-progenitor cluster. Scale bars: 100 μm.

Article Snippet: Cells were stimulated with recombinant mouse IGF1 (Cat No. 791-MG-050, R&D systems) for 0, 30, 60, or 180 min.

Techniques: RNAscope, In Situ Hybridization, Expressing

Journal: REPRODUCTION

Article Title: Increased apoptosis in bovine blastocysts exposed to high levels of IGF1 is not associated with downregulation of the IGF1 receptor

doi: 10.1530/rep-10-0336

Figure Lengend Snippet: Figure 1 Effect of different IGF1 concentrations on the percentage of in vitro-produced blastocysts displaying different inner cell mass (ICM)/total cell number (TCN) proportions. Within ICM/TCN categories, bars with different letters differ significantly (P%0.05).

Article Snippet: For the IGF1 treatments, a vial containing 50 mg of lyophilised recombinant human IGF1 (291-G1, R&D systems, Wiesbaden-Norderstadt, Germany) was rehydrated with 500 ml of 0.1% PBS/BSA according to the manufacturer’s recommendations and stored at K20 8C in 10 ml aliquots (1000 ng).

Techniques: In Vitro, Produced

Journal: REPRODUCTION

Article Title: Increased apoptosis in bovine blastocysts exposed to high levels of IGF1 is not associated with downregulation of the IGF1 receptor

doi: 10.1530/rep-10-0336

Figure Lengend Snippet: Figure 2 Relative transcript abundance (meanGS.E.M.) of develop- mentally important genes in day-8 blastocysts cultured in the presence of different IGF1 concentrations from the zygote stage onwards. Bars with different superscripts within each gene transcript indicate a significant difference (P%0.05).

Article Snippet: For the IGF1 treatments, a vial containing 50 mg of lyophilised recombinant human IGF1 (291-G1, R&D systems, Wiesbaden-Norderstadt, Germany) was rehydrated with 500 ml of 0.1% PBS/BSA according to the manufacturer’s recommendations and stored at K20 8C in 10 ml aliquots (1000 ng).

Techniques: Cell Culture

Journal: REPRODUCTION

Article Title: Increased apoptosis in bovine blastocysts exposed to high levels of IGF1 is not associated with downregulation of the IGF1 receptor

doi: 10.1530/rep-10-0336

Figure Lengend Snippet: Figure 3 Three-dimensional reconstruction of confocal laser microscopy images showing protein localisation of the IGF1R and TP53 in day- 8 blastocysts cultured in the presence of different concentrations of IGF1. PI, propidium iodide. Note the lower expression of the IGF1R in the inner cell mass of the control group compared with the IGF1 groups and the lower expression of TP53 in the trophectoderm in the supraphysiolo- gical IGF1 group (1000 ng/ml).

Article Snippet: For the IGF1 treatments, a vial containing 50 mg of lyophilised recombinant human IGF1 (291-G1, R&D systems, Wiesbaden-Norderstadt, Germany) was rehydrated with 500 ml of 0.1% PBS/BSA according to the manufacturer’s recommendations and stored at K20 8C in 10 ml aliquots (1000 ng).

Techniques: Microscopy, Cell Culture, Expressing, Control

Journal: REPRODUCTION

Article Title: Increased apoptosis in bovine blastocysts exposed to high levels of IGF1 is not associated with downregulation of the IGF1 receptor

doi: 10.1530/rep-10-0336

Figure Lengend Snippet: Figure 5 Relative signal strength (RSS) values (meanGS.E.M.) of TP53 in day-8 blastocysts treated with different concentrations of IGF1. ICM, inner cell mass; TE, trophectoderm. Between groups, bars with different superscripts indicate a significant difference (TE: a, b; total RSS: A, B; P%0.05). Within groups, the asterisk indicates significant differences between the ICM and the TE (P%0.05).

Article Snippet: For the IGF1 treatments, a vial containing 50 mg of lyophilised recombinant human IGF1 (291-G1, R&D systems, Wiesbaden-Norderstadt, Germany) was rehydrated with 500 ml of 0.1% PBS/BSA according to the manufacturer’s recommendations and stored at K20 8C in 10 ml aliquots (1000 ng).

Techniques:

Journal: REPRODUCTION

Article Title: Increased apoptosis in bovine blastocysts exposed to high levels of IGF1 is not associated with downregulation of the IGF1 receptor

doi: 10.1530/rep-10-0336

Figure Lengend Snippet: Figure 4 Relative signal strength (RSS) values (meanGS.E.M.) of the IGF1R in day-8 blastocysts treated with different concentrations of IGF1 and displaying immunofluorescence in the inner cell mass (ICM) and the trophectoderm (TE). Between groups, bars with different super- scripts indicate a significant difference (ICM: a, b; TE: c, d; total RSS: A, B; P%0.05). Within groups, the asterisk indicates significant differences between the ICM and the TE (P%0.05).

Article Snippet: For the IGF1 treatments, a vial containing 50 mg of lyophilised recombinant human IGF1 (291-G1, R&D systems, Wiesbaden-Norderstadt, Germany) was rehydrated with 500 ml of 0.1% PBS/BSA according to the manufacturer’s recommendations and stored at K20 8C in 10 ml aliquots (1000 ng).

Techniques:

Journal: Journal of Molecular Endocrinology

Article Title: Human aortic smooth muscle cells are insulin resistant at the receptor level but sensitive to IGF1 and IGF2

doi: 10.1677/jme-09-0021

Figure Lengend Snippet: Figure 1 Expression of IR and IGF1R mRNA and proteins in HASMC. The relative amount of IGF1R to IR mRNA expression was measured by real-time RT-PCR, where IR was set to 1. The IGF1R and IR protein levels were analyzed by ELISA and are expressed as amount receptor protein relative to amount of total protein, IR was set to 1. The IGF1R to IR mRNA ratio (5.3G1.3) and the IGF1R to IR protein ratio (5.5G1.2) was found to be almost identical. ***Denotes P!0.001 (meanGS.E.M., nZ4).

Article Snippet: After washing away the unbound fraction with PBS–T, the secondary antibody was added and incubated on a shaker at RT for 2 h. The biotinylated anti-human IGF1R antibody (BAF 391; R&D Systems) was diluted as 0.5 mg/l in 0.1 w/v % HSA in PBS.

Techniques: Expressing, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay

Journal: Journal of Molecular Endocrinology

Article Title: Human aortic smooth muscle cells are insulin resistant at the receptor level but sensitive to IGF1 and IGF2

doi: 10.1677/jme-09-0021

Figure Lengend Snippet: Figure 3 Detection of IR and IGF1R b-subunits in HASMC. IR and IGF1R b-subunit protein and evidence for the presence of hybrid IR/IGF1R were demonstrated by immunoprecipitation and western blot. Insulin receptor (IR) b-subunit (95 kDa) and IGF1R (IGF1R) b-subunit (97 kDa) were detected by western blot on the same membrane implying co-precipitation. Figures show blots after immunoprecipitation (IP) against either IR (left column, A and B), or IGF1R (right column, C and D) and immunoblot against the IR b-subunit (A and D) and the IGF1R b-subunit (B and C). Similar results were obtained in four independent experiments.

Article Snippet: After washing away the unbound fraction with PBS–T, the secondary antibody was added and incubated on a shaker at RT for 2 h. The biotinylated anti-human IGF1R antibody (BAF 391; R&D Systems) was diluted as 0.5 mg/l in 0.1 w/v % HSA in PBS.

Techniques: Immunoprecipitation, Western Blot, Membrane

Journal: Journal of Molecular Endocrinology

Article Title: Human aortic smooth muscle cells are insulin resistant at the receptor level but sensitive to IGF1 and IGF2

doi: 10.1677/jme-09-0021

Figure Lengend Snippet: Figure 4 (A) Activation of IR b-subunit in HASMC. Effect of insulin (dark gray bars), IGF1 (black bars), and IGF2 (gray bars) on tyrosine phosphorylation of IR b-subunit were analyzed by immunoprecipitation of IR by an anti-IR b-subunit antibody and immunoblotting with an anti-phosphotyrosine antibody (PY20) after exposure of near confluent HASMC to insulin, IGF1, and IGF2 at the concentrations of 10K10–10K8 mol/l for 10 min. Data are means from four independent experiments. Data were quantified by densitometry and are expressed as phosphor- ylated/total protein, fractional of maximal response. *P!0.05. (B) Activation of IGF1R b-subunit in HASMC. Effects of insulin (dark gray bars), IGF1 (black bars), and IGF2 (gray bars) on tyrosine phosphorylation of IGF1R b-subunit were analyzed by immunoprecipitation of IGF1R by an anti IGF1R b-subunit antibody and immunoblotting with an anti-phosphotyrosine antibody (PY20) after exposure of near confluent HASMC to insulin, IGF1, and IGF2 at the concentrations of 10K10– 10K8 mol/l for 10 min. Data are means from four independent experiments. Data were quantified by densitometry and are expressed as phosphorylated/total protein, fractional of maximal response. *P!0.05.

Article Snippet: After washing away the unbound fraction with PBS–T, the secondary antibody was added and incubated on a shaker at RT for 2 h. The biotinylated anti-human IGF1R antibody (BAF 391; R&D Systems) was diluted as 0.5 mg/l in 0.1 w/v % HSA in PBS.

Techniques: Activation Assay, Phospho-proteomics, Immunoprecipitation, Western Blot

Journal: Journal of Molecular Endocrinology

Article Title: Human aortic smooth muscle cells are insulin resistant at the receptor level but sensitive to IGF1 and IGF2

doi: 10.1677/jme-09-0021

Figure Lengend Snippet: Figure 8 Proposed model for insulin resistance in HASMC. The insulin resistance of vascular smooth muscle is located at the receptor level. Due to the low number of IRs there are not enough receptors activated by insulin to generate a downstream signal. However, the IR b-subunit is activated by IGF1 when incorporated into hybrid IR/IGF1R. Both IGF1 and IGF2 activate the IGF1R and the hybrid IR/IGF1R and this elicits downstream signaling and biological effects.

Article Snippet: After washing away the unbound fraction with PBS–T, the secondary antibody was added and incubated on a shaker at RT for 2 h. The biotinylated anti-human IGF1R antibody (BAF 391; R&D Systems) was diluted as 0.5 mg/l in 0.1 w/v % HSA in PBS.

Techniques:

Journal: Frontiers in Oncology

Article Title: Growth hormone receptor antagonism downregulates ATP-binding cassette transporters contributing to improved drug efficacy against melanoma and hepatocarcinoma in vivo

doi: 10.3389/fonc.2022.936145

Figure Lengend Snippet: GHRA suppresses syngeneic mouse melanoma growth in vivo . (A) Mouse Fluc-B16-F10 cells grafted intradermally on the right flank of syngeneic C57BL6/J wild-type (WT) and GHA male mice (transgenic for bGH G119K GHR antagonist) (n=6). Tumor growth over 4-weeks was followed by luminescent imaging following luciferin injections. Luciferin signal was quantified and plotted on the right. The changes in tumor volume (Fluc-B16-F10 in WT and GHA mice) from digital caliper measurement (B) and tumor mass (C) corroborate the suppressed tumor growth in GHA mice which has markedly lower serum IGF1 levels (D) due to presence of a circulating GHRA. Western-blot analysis of the GH downstream signaling mediators – phosphorylated STAT5, AKT and SRC kinase in the tumors of GHA and WT mice (E) . B16-F10 cells in culture when treated for 72-hours with serum collected from WT and GHA mice showed suppressed growth rate in the GHA mouse serum (F) . (*p < 0.05, mouse studies – repeated measure using SPSS; cell viability - Students t test, n = 3).

Article Snippet: The sera were then collected and measured by ELISA for IGF1 (#22-IG1MS-E01, ALPCO, Salem, NH, USA) according to the manufacture’s guidelines.

Techniques: In Vivo, Transgenic Assay, Imaging, Western Blot

Journal: Frontiers in Oncology

Article Title: Growth hormone receptor antagonism downregulates ATP-binding cassette transporters contributing to improved drug efficacy against melanoma and hepatocarcinoma in vivo

doi: 10.3389/fonc.2022.936145

Figure Lengend Snippet: Effect of GHRA on response of syngeneic mouse hepatocellular carcinoma tumors to sorafenib treatment in vivo. (A) Mouse Hepa1-6 hepatocellular carcinoma (HCC) cells respond to bovine GH stimulation in culture showing increased cell viability and STAT5 activation after 72 hours (A) as well as increase in the EC50 dose of sorafenib (B) . Additionally, GH treatment increased ABC transporter levels in the cultured Hepa1-6 cells (C) . Mouse Hepa1-6 cells grafted subcutaneously on the right flank of syngeneic C57BL6/J wild-type (WT) and GHA mice (transgenic for bGH G119K GHR antagonist) (n=6). The changes in tumor volume (Hepa1-6 in WT and GHA mice) from digital caliper measurement (D) and post-dissection tumor mass (E) show suppressed tumor growth and improved tumoral response to sorafenib in the GHA mice. (F) Spearman correlation analysis for transcript levels of GHR and ABC transporter and IGF1R and ABC transporters in the tumor of 371 HCC patients in the TCGA cohort. (G) Serum IGF1 levels of Hepa1-6 tumor-bearing GHA mice are significantly lower than that of tumor-bearing WT mice. (H–J) Correlation of overall survival probability of 371 HCC patients in the TCGA cohort with RNA expression of IGF1R (H) , or ABCB1 (I) , or ABCC1 (J) . (*p < 0.05, ***p < 0.001, mouse studies – repeated measure using SPSS; other assays - Students t test, n = 3).

Article Snippet: The sera were then collected and measured by ELISA for IGF1 (#22-IG1MS-E01, ALPCO, Salem, NH, USA) according to the manufacture’s guidelines.

Techniques: In Vivo, Activation Assay, Cell Culture, Transgenic Assay, Dissection, RNA Expression

Journal: Scientific Reports

Article Title: Insulin-like growth factor 2 and autophagy gene expression alteration arise as potential biomarkers in Parkinson’s disease

doi: 10.1038/s41598-022-05941-1

Figure Lengend Snippet: Plasma IGF1 and IGF2 levels in PD patients do not correlate with motor scores of PD patients. ( A ) Pearson correlation analysis between IGF2 levels in PD patients with H&Y (top panel, r = 0.1371), UPDRSIII (middle panel, r = − 0.07482) scores and evolution disease time (bottom panel, r = − 0.1951). ( B ) Correlation analysis between IGF1 levels in PD patients with H&Y (top panel, r = 0.1498), UPDRSIII (middle panel r = − 0.07823) scores and evolution disease time (bottom panel, r = − 0.09742).

Article Snippet: The plasma levels of IGF1 and IGF2 were measured by ELISA (ALPCO, 22-IGFHU-E01, and 22-IG2HU-E01) according to the manufacturer’s instructions.

Techniques: Clinical Proteomics