total igf- 1  (ALPCO)

Journal: Journal of Feline Medicine and Surgery

Article Title: Signalment, clinicopathological findings, management practices and comorbidities in cats with diabetes mellitus in Germany: cross-sectional study of 144 cases

doi: 10.1177/1098612X241303303

Figure Lengend Snippet: Comparison of sex, age, fructosamine concentration and insulin dose between diabetic cats with and without increased IGF-1 (IGF-1 ⩽746 ng/ml) suspicious for hypersomatotropism

Article Snippet: Veterinary practices and owners of diabetic cats could participate if they submitted a completed questionnaire (see Appendix 1 in the supplementary material ) alongside blood samples for haematology (ADVIA 2120i; Siemens Diagnostics), serum biochemistry, DGGR ester lipase activity, concentration of fructosamine, b-hydroxybutyrate (BHB) (all measured by AU 5800, AU 680; Beckman Coulter and Alinity CI-Systems), total thyroxine (TT4), COB and IGF-1 (all three measured using IMMULITE 2000 XPi Immunoassay System; Siemens Medical Solutions Diagnostics).

Techniques: Comparison, Concentration Assay

Journal: Frontiers in Oncology

Article Title: IGF1R inhibition and PD-1 blockade improve anti-tumor immune response in epithelial ovarian cancer

doi: 10.3389/fonc.2024.1410447

Figure Lengend Snippet: DC treated with IGF1R inhibitors affect OC cell line proliferation. Human leukemic THP-1 cells were differentiated to DC by adding 40 ng/ml IL-4, 20 ng/ml GM-CSF, 20 ng/ml TNFα and 200 ng/ml ionomycin for 48 h and 5 μM of AEW for 48 h. Differentiated DC, IGF1R-treated-DC, THP-1 or IGF1R-treated-THP-1 were each co-cultured with CFSE-pre-labeled OC cell lines for 48 h. Representative flow cytometry histograms of CFSE proliferation assays for (A) ES2 cells (left) and flow cytometry analysis for ES2 cells (right). (B) CFSE dilution assay for SKOV3 cell proliferation (left) and flow cytometry analysis for SKOV3 cells (right). (C) CFSE dilution assay for Ku cell proliferation (left) and flow cytometric analysis for Ku cell (right). The graphs represent OC cell lines CFSE MFI from three independent experiments. * p<0.05. Bars represent SEM values.

Article Snippet: Cells were then washed and incubated with primary antibodies against total IGF1R (sc-81,167, Santa Cruz Biotechnology, Inc.) and phospho IGF1R (Y1135/1136, Cell Signaling) for 1 h at RT.

Techniques: Cell Culture, Labeling, Flow Cytometry, Dilution Assay

Journal: Frontiers in Oncology

Article Title: IGF1R inhibition and PD-1 blockade improve anti-tumor immune response in epithelial ovarian cancer

doi: 10.3389/fonc.2024.1410447

Figure Lengend Snippet: Treatment regimen and the presentation of ID8 ovarian cancer. 32 C57BL/6 mice were inoculated intraperitoneally with 3.5 *10 6 ID8-mCherry cells, and randomly divided into 4 groups. 14 days after tumor cell injection, mice were treated with either dilution buffer as control, 50 mg/kg IGF1R inhibitor daily for a week, 200 µg anti-PD-1 twice per week for 2 weeks, or anti-PD-1/IGF1R. (A) Schematic regimen for single and combined treatment. (B) Graph representing the mean tumor weight in the single treatments and the combined treatment. (C) The survival of tumor-bearing mice was monitored by Kaplan-Meier analysis and statistical analyses were performed with Log rank test. (D) Graph of the mean survival time. Bars represent mean ± SEM.* p < 0.05.

Article Snippet: Cells were then washed and incubated with primary antibodies against total IGF1R (sc-81,167, Santa Cruz Biotechnology, Inc.) and phospho IGF1R (Y1135/1136, Cell Signaling) for 1 h at RT.

Techniques: Injection, Control

Journal: Frontiers in Oncology

Article Title: IGF1R inhibition and PD-1 blockade improve anti-tumor immune response in epithelial ovarian cancer

doi: 10.3389/fonc.2024.1410447

Figure Lengend Snippet: Co-inhibition of PD-1 and IGF1R increased CD8 T-cell populations in the ovarian cancer microenvironment. Three tumors from each group were resected and stained with CD45+, CD4+ and CD8a+ markers. The mice were treated with either dilution buffer as control, 50 mg/kg IGF1R inhibitor daily for a week, 200 µg anti-PD-1 twice per week for 2 weeks, or anti-PD-1/IGF1R. The prevalence of (A) leukocyte (CD45+), (B) T-helper (CD4+) and (C) T-cytotoxic (CD8a+) in single cell suspensions from ID8 tumor-bearing mice are shown. Data were obtained from FCM assays, with 3 mice/group. All cell subtypes were CD45+ gated. Bars represent mean ± SEM.* p < 0.05.

Article Snippet: Cells were then washed and incubated with primary antibodies against total IGF1R (sc-81,167, Santa Cruz Biotechnology, Inc.) and phospho IGF1R (Y1135/1136, Cell Signaling) for 1 h at RT.

Techniques: Inhibition, Staining, Control

Journal: Frontiers in Oncology

Article Title: IGF1R inhibition and PD-1 blockade improve anti-tumor immune response in epithelial ovarian cancer

doi: 10.3389/fonc.2024.1410447

Figure Lengend Snippet: Co-inhibition of PD-1 and IGF1R increase DC prevalence in the ovarian cancer microenvironment. Mice were inoculated intraperitoneally with 3.5*106 ID8 tumor cells, mice were treated with either dilution buffer as control, 50 mg/kg IGF1R inhibitor daily for a week, 200 µg anti-PD-1 twice per week for 2 weeks or anti-PD-1/IGF1R. Three tumors from each group were harvested and stained with CD45+, CD11c+ and CD86+, CD8a+ and CD11b markers. DC (CD45+CD11c+), and classic dendritic cells (cDC1 and cDC2) were quantified in cells isolated from tumors. The rates of (A) CD11c+ DC (left) and histogram of CD11c DC expression on CD45+ cells (right), (B) CD86 positive CD11c DCs, (C) cDC1 (CD11c+CD86+CD8a+CD11b-) and (D) cDC2 (CD11c+CD86+CD11b+) isolated from ID8 tumor-bearing mice, were measured and analyzed using FCM assays, 3 mice/group. All cell subtypes were gated on CD45+. Bars represent mean ± SEM. * p < 0.05.

Article Snippet: Cells were then washed and incubated with primary antibodies against total IGF1R (sc-81,167, Santa Cruz Biotechnology, Inc.) and phospho IGF1R (Y1135/1136, Cell Signaling) for 1 h at RT.

Techniques: Inhibition, Control, Staining, Isolation, Expressing

Journal: Frontiers in Oncology

Article Title: IGF1R inhibition and PD-1 blockade improve anti-tumor immune response in epithelial ovarian cancer

doi: 10.3389/fonc.2024.1410447

Figure Lengend Snippet: Exploring the effect of combined anti-PD-1/IGF1R treatment on the tumor microenvironment, through RNA-seq. Mice were inoculated intraperitoneally with 3.5*106 ID8 ovarian tumor cells. Tumors were harvested from 2 mice in the control group, 4 mice in the IGF1R inhibitor-treated group, 3 mice in the anti-PD-1-treated group, and 4 mice in the anti-PD-1/IGF1R-treated group. Subsequently, RNA-seq was performed using RNA extracted from the 13 tumors. (A) A hierarchical clustering heatmap of the 4 experimental groups based on gene expression RNA-seq. (B) A Venn diagram showing the common and unique differentially expressed genes in the combined anti-PD-1/IGF1R treatment group compared to control, IGF1R inhibitor and ant-PD-1 treatments. Target genes were filtered by a fold change >1.5 or < -1.5. (C) GO analysis bar graph of the 443 shared genes, showing the enriched biological processes.

Article Snippet: Cells were then washed and incubated with primary antibodies against total IGF1R (sc-81,167, Santa Cruz Biotechnology, Inc.) and phospho IGF1R (Y1135/1136, Cell Signaling) for 1 h at RT.

Techniques: RNA Sequencing, Control, Gene Expression

Journal: Frontiers in Oncology

Article Title: IGF1R inhibition and PD-1 blockade improve anti-tumor immune response in epithelial ovarian cancer

doi: 10.3389/fonc.2024.1410447

Figure Lengend Snippet: Summary of pair-wise differential expression analysis.

Article Snippet: Cells were then washed and incubated with primary antibodies against total IGF1R (sc-81,167, Santa Cruz Biotechnology, Inc.) and phospho IGF1R (Y1135/1136, Cell Signaling) for 1 h at RT.

Techniques: Quantitative Proteomics, Control

Journal: Frontiers in Oncology

Article Title: IGF1R inhibition and PD-1 blockade improve anti-tumor immune response in epithelial ovarian cancer

doi: 10.3389/fonc.2024.1410447

Figure Lengend Snippet: Up- or down-regulated target genes significantly affected by anti-PD1/IGF1R treatment. Mice were inoculated intraperitoneally with 3.5*106 ID8 ovarian tumor cells (n=8/group). Tumors were harvested from 2 mice in the control group, 4 mice in the IGF1R inhibitor-treated group, 3 mice in the anti-PD-1-treated group, and 4 mice in the anti-PD-1/IGF1R-treated group. Subsequently, RNA-seq was performed using RNA extracted from the 13 tumors. Volcano plots showing expression fold changes (X-axis) versus p value (Y-axis). Significantly up-regulated genes are presented in green on the volcano’s right side and the significantly down-regulated genes are presented in red on the left side of the volcano. (A) anti-PD-1/IGF1R treatment compared to Control. (B) anti-PD-1/IGF1R treatment compared to Anti-PD-1 treatment. (C) anti-PD-1/IGF1R treatment compared to IGF1R inhibitor treatment.

Article Snippet: Cells were then washed and incubated with primary antibodies against total IGF1R (sc-81,167, Santa Cruz Biotechnology, Inc.) and phospho IGF1R (Y1135/1136, Cell Signaling) for 1 h at RT.

Techniques: Control, RNA Sequencing, Expressing

Journal: The FASEB Journal

Article Title: Impaired myoblast differentiation and muscle IGF‐1 receptor signaling pathway activation after N‐glycosylation inhibition

doi: 10.1096/fj.202400213rr

Figure Lengend Snippet: FIGURE 4 IGF-1 production and IGF1R signaling pathway activation in TA muscle of WT and MLC/mIgf-1 mice treatment with low TUN dose (0.1 mg/kg of TUN for 15 days). (A) Expression of endogenous Igf-1 mRNA isoforms (i.e., Igf-1Ea and Igf-1Ec mRNAs) and total Igf-1 (i.e., endogenous and mIgf-1 transgene mRNAs) in the TA muscle of WT (n = 3) and MLC/mIgf-1 (n = 3) mice treated with TUN. (B) Representative immunoblotting and (C) band densitometry analysis of TA muscle of WT and MLC/mIgf1 mice using an antibody directed against mature mouse IGF-1 sequence. Three bands at a molecular weight of ~22, ~17, and ~12 kDa were detected in TA muscle of MLC/ mIgf1 mice. Recombinant mouse IGF-1 mature protein was loaded as a positive control for mature IGF-1 (~7 kDa). (D) Representative immunoblotting and (E) band densitometry analysis of TA muscle of WT and MLC/mIgf1 mice using antibodies directed against phosphorylated (pIGF1R) and total IGF1R, phosphorylated (pAKT) and total AKT, and phosphorylated (pERK1/2) and total ERK1/2. All bar charts are presented as mean values ± SD. Significant differences were determined using unpaired t-test (C) or two-way ANOVA followed by Tukey's multiple comparison post hoc tests (A, E). *Significantly different compared to CTR; #Significantly different compared to WT mice; * and # p ≤ .05; ** p ≤ .01 *** and ### p ≤ .001.

Article Snippet: An antibody against total IGF1R β (1:1000; cat. n. 3027 Cell Signaling Technology) was incubated at room temperature for 1 h followed by incubation at room temperature for 1 h with 1:200 Rhodamine RedTM- X (RRX) AffiniPureTM Goat Anti- Rabbit IgG (H+L) (Jackson ImmunoResearch Europe Ltd.).

Techniques: Activation Assay, Expressing, Western Blot, Sequencing, Molecular Weight, Recombinant, Positive Control, Comparison

Journal: The FASEB Journal

Article Title: Impaired myoblast differentiation and muscle IGF‐1 receptor signaling pathway activation after N‐glycosylation inhibition

doi: 10.1096/fj.202400213rr

Figure Lengend Snippet: FIGURE 5 Effect of TUN treatment on IGF1R production and IGF1R signaling pathway activation in C2C12. (A) IGF1R and IGF1R proreceptor production in C2C12 cells treated with 0.01 μg/mL of TUN and harvested at day 1. (B) Immunofluorescence analysis of C2C12 cells stained with ant-IGF1R and DAPI, the scale bar represents 200 μm. (C) Representative immunoblotting and (D) band densitometry analysis of IGF-1-induced IGF-1R signaling pathway activation in CTR- and TUN-treated C2C12 cells using antibodies directed against phosphorylated (pIGF1R) and total IGF1R, phosphorylated (pAKT) and total AKT, and phosphorylated (pERK1/2) and total ERK1/2. The calculation of pIGF1R level was normalized against total lysed protein instead of total IGF1R to account for the marked reduction of total IGF1R after TUN treatment. All bar charts are presented as mean values ± SD. Significant differences were determined using unpaired t-test (A) or one-way ANOVA followed by Tukey's multiple comparison post hoc tests (D). *Significantly different compared to CTR, #Significantly different compared to IGF-1-treated cells; ** p ≤ .01, *** p ≤ .001; ## p ≤ .01, ### p ≤ .001.

Article Snippet: An antibody against total IGF1R β (1:1000; cat. n. 3027 Cell Signaling Technology) was incubated at room temperature for 1 h followed by incubation at room temperature for 1 h with 1:200 Rhodamine RedTM- X (RRX) AffiniPureTM Goat Anti- Rabbit IgG (H+L) (Jackson ImmunoResearch Europe Ltd.).

Techniques: Activation Assay, Immunofluorescence, Staining, Western Blot, Comparison