recombinant mouse igf1 protein  (MedChemExpress)

Journal: BMC Medicine

Article Title: IGF1-mediated mesenchymal-endothelial transition as a potential regulatory target in calcific aortic valve disease

doi: 10.1186/s12916-025-04433-z

Figure Lengend Snippet: IGF1 identified as a key molecule mediating MEndT. A Conduct protein-protein interaction network analysis (PPI) on the overlapping differentially expressed genes between pVECs vs. pVICs and pVICs-OM8d vs. pVICs. B Analyze the FPKM values of SOD2, CCL2, CCN2, IGF1, and COL3A1 in the transcriptome sequencing of pVICs-OM8d vs. pVICs, normalized to the control pVICs group. Values are mean ± SD of 4 biological replicates. Statistical tests used: ANOVA. C , D WB ( C ) and qPCR ( D ) were used to detect the protein expression of IGF1, IGF1R, P-IGF1R, and the mRNA expression levels of IGF1, IGF1R in pVICs cultured for 8 days in GM or OM. D Normalized to pVICs and GAPDH. Values are mean ± SD of 3 independent experiments. Statistical tests used: ANOVA. E , F Statistical analysis of tube formation assays of pVICs with exogenous addition of IGF1 and BMS-536924 during OM induction ( E ), and after knockdown of IGF1 expression. Normalized to GM group ( F ). Normalized to pVICs. Values are mean ± SD. 3 biological replicates, with 3 random measurements within each replicate, n =9. Statistical tests used: ANOVA. G , H Assessed the changes in protein expression ( G ) of CDH5, CD31, α-SMA, PI3K, Akt, P-Akt, and HIF-1α in pVICs after the exogenous addition of recombinant IGF1 and inhibitor BMS-536924 in GM and OM, and changes in mRNA expression levels ( H ) of CDH5, CD31, α-SMA. H Normalized to pVICs and GAPDH. Values are mean ± SD of 3 independent experiments. Statistical tests used: ANOVA. I , J Detect the protein ( J ) and mRNA ( I ) expression of CDH5, CD31, α-SMA in pVICs after knockdown of IGF1 expression during OM induction. I Normalized to pVICs and GAPDH. Values are mean ± SD of 3 independent experiments. Statistical tests used: ANOVA. K , L Tube formation assays in pVICs with exogenous addition of IGF1 and BMS-536924 during OM induction, and after knockdown of IGF1 expression, scale bars: 200 µm

Article Snippet: Each group received intraperitoneal injections every 2 days with either saline (100 μl), recombinant mouse IGF1 protein (1 μM, 100 μl; MCE, HY-P7070), or BMS-536924 (1 mg; MCE, HY-10262).

Techniques: Sequencing, Control, Expressing, Cell Culture, Knockdown, Recombinant

Journal: BMC Medicine

Article Title: IGF1-mediated mesenchymal-endothelial transition as a potential regulatory target in calcific aortic valve disease

doi: 10.1186/s12916-025-04433-z

Figure Lengend Snippet: IGF1-mediated MEndT and disease progression in the AVWI mouse mode. A Schematic diagram of the animal experiment procedure. B Echocardiographic evaluation of the sham surgery group and the AVWI mouse groups with intraperitoneal injection of saline, IGF1, or BMS-536924. Parameters assessed included: aortic valve annulus diameter (mm) and transaortic peak velocity (mm/s). C , D Statistical analysis of the aortic valve annulus diameter (mm) and transaortic peak velocity (mm/s) in each group of mice. n =8. Values are mean ± SD. Statistical tests used: ANOVA. E HE staining (scale bars: 200 µm) and multiplex immunofluorescence histology (scale bars: 100 µm, 50 µm, or 20 µm) for CD31, tdTomato, and DAPI of mouse aortic valve paraffin sections. Yellow arrows indicate CD31-positive cells labeled with tdTomato. F , G Statistical analysis of the aortic valve thickness (µm) (HE staining, E ) and tdTomato labeled cells expressing CD31 within the aortic valve region (immunofluorescence staining, G ) in each group of mice. n =8. Values are mean ± SD. Statistical tests used: ANOVA

Article Snippet: Each group received intraperitoneal injections every 2 days with either saline (100 μl), recombinant mouse IGF1 protein (1 μM, 100 μl; MCE, HY-P7070), or BMS-536924 (1 mg; MCE, HY-10262).

Techniques: Biomarker Discovery, Injection, Saline, Staining, Multiplex Assay, Immunofluorescence, Labeling, Expressing

Journal: Cell Communication and Signaling : CCS

Article Title: Megakaryocytic IGF1 coordinates activation and ferroptosis to safeguard hematopoietic stem cell regeneration after radiation injury

doi: 10.1186/s12964-024-01651-5

Figure Lengend Snippet: MKs become a predominant component and IGF1 source of HSC niche after radiation injury. ( A and B ) The number and frequency of MKs in the BM of mice at indicated time post IR ( n = 6). ( C ) Schematic illustration of MK subpopulation assay. ( D ) Heatmap showing the expression of genes related to each MK subpopulation at indicated time post IR. ( E ) Radar plot showing functional shift of MKs at indicated time post IR. ( F ) Flow cytometric analysis of the fraction of MK subpopulations in the BM of mice at indicated time post IR ( n = 6). ( G ) Immunostaining analysis of positional relationship between HSCs and MKs in the BM of mice at 3 dpi. The yellow arrow indicates HSC. The dashed line outlines MK. Scale bar, 10 μm ( n = 60). ( H ) Heatmap showing the expression of megakaryocytic secretory factors at indicated time post IR. ( I ) Reactome pathway enrichment analysis of BM MKs at 3 dpi. ( J ) Flow cytometric analysis of IGF1 expression in BM MKs of mice at indicated time post IR ( n = 6). ( K ) Relative IGF1 levels in the BM fluid of mice at indicated time post IR ( n = 6). ( L ) Flow cytometric analysis of IGF1 expression in BMC and platelets of mice at 3 dpi. Mono, monocyte; Mac, macrophage; Dc, dendritic cell; Endo, endothelium; MSC, mesenchymal stromal cell. ( n = 6). ( M ) Violin plots showing Igf1 expression in different cell clusters at homeostasis. Hematopoietic stem and progenitor cell, HSPC; osteoblast, OB. ( N ) MK numbers in the BM of WT and Mpl hlb219 mice at 3 dpi ( n = 6). ( O ) Relative IGF1 levels in the BM fluid of WT and Mpl hlb219 mice at 3 dpi ( n = 6). Data represent mean ± SD. * P < 0.05, ** P < 0.01, NS: no significance. Two-tailed unpaired student’s t -test unless stated otherwise. One-way ANOVA was used for calculating P values in ( F ), ( N ) and ( O )

Article Snippet: For IGF1 administration, mice were subcutaneously treated with a dose of 200 µg/kg recombinant mouse IGF1 (R&D Systems, Minneapolis, MN, USA).

Techniques: Expressing, Functional Assay, Immunostaining, Two Tailed Test

Journal: Cell Communication and Signaling : CCS

Article Title: Megakaryocytic IGF1 coordinates activation and ferroptosis to safeguard hematopoietic stem cell regeneration after radiation injury

doi: 10.1186/s12964-024-01651-5

Figure Lengend Snippet: Megakaryocytic IGF1 promotes functional expansion of HSCs after radiation injury. ( A ) Flow cytometric analysis of p-IGF1R expression in HSCs of mice at day 1 post IGF1 administration ( n = 6). ( B ) Flow cytometric gating strategy, frequency and the number of HSCs in the BM of mice at day 1 post IGF1 administration ( n = 6). ( C and D ) Experimental design and PB chimerism at 16 weeks post-competitive transplantation of HSCs from mice at day 1 post IGF1 administration ( n = 6). ( E ) Frequencies of HSCs in the ex vivo culture system with different concentrations of IGF1 at day 10 ( n = 6). ( F ) Colony numbers of per 10 3 lineage – cells sorted from HSC cultures as indicated ( n = 6). ( G ) Experimental design of an ex vivo co-culture experiment. ( H ) Relative IGF1 contents in the supernatant of the indicated culture ( n = 6). ( I ) Frequencies of HSCs and colony numbers of per 10 3 lineage – cells sorted from various HSC cultures. 10 µM AG1024 was added in the culture in the indicated group ( n = 6). Data represent mean ± SD. * P < 0.05, ** P < 0.01, NS: no significance. One-way ANOVA unless stated otherwise. Two-tailed unpaired student’s t -test was used for calculating P values in ( A ), ( B ) and ( D )

Article Snippet: For IGF1 administration, mice were subcutaneously treated with a dose of 200 µg/kg recombinant mouse IGF1 (R&D Systems, Minneapolis, MN, USA).

Techniques: Functional Assay, Expressing, Transplantation Assay, Ex Vivo, Co-Culture Assay, Two Tailed Test

Journal: Cell Communication and Signaling : CCS

Article Title: Megakaryocytic IGF1 coordinates activation and ferroptosis to safeguard hematopoietic stem cell regeneration after radiation injury

doi: 10.1186/s12964-024-01651-5

Figure Lengend Snippet: Punctual IGF1 administration effectively mitigates myelosuppression induced by radiation injury. ( A and B ) Flow cytometric analysis of expression of p-IGF1R and p-mTOR in HSCs in the BM of mice with or without IGF1 supplementation at 1dpi ( n = 6). ( C ) Flow cytometric analysis of cell cycle of HSCs in the BM of mice with or without IGF1 supplementation at 1dpi ( n = 6). ( D ) Flow cytometric analysis of GFP-LC3 expression in HSCs in the BM of GFP-LC3 mice with or without IGF1 supplementation at 1 dpi ( n = 6). ( E ) Flow cytometric analysis of Ferritin and FerroOrange of HSCs in the BM of mice with or without IGF1 supplementation at 1 dpi ( n = 6). ( F and G ) Flow cytometric analysis cell death and lipid peroxidation of HSCs in the BM of mice with or without IGF1 supplementation at 1dpi ( n = 6). ( H ) The number of HSCs in the BM of mice with or without IGF1 supplementation at indicated time post IR ( n = 6). ( I ) WBC, RBC, and platelet counts in the PB of mice with or without IGF1 supplementation at indicated time post IR ( n = 6). ( J ) Schematic illustration of the protective role of megakaryocytic IGF1 in safeguarding HSC regeneration. Data represent mean ± SD. * P < 0.05, ** P < 0.01. Two-tailed unpaired student’s t -test

Article Snippet: For IGF1 administration, mice were subcutaneously treated with a dose of 200 µg/kg recombinant mouse IGF1 (R&D Systems, Minneapolis, MN, USA).

Techniques: Expressing, Two Tailed Test

Journal: iScience

Article Title: Contrasting consequences of podocyte insulin-like growth factor 1 receptor inhibition

doi: 10.1016/j.isci.2024.109749

Figure Lengend Snippet: In vitro differential suppression of podocyte IGF1R activity reveals that partial inhibition is beneficial but near total loss is highly detrimental (A) Representative Western blot shows >90% reduction of IGF1R protein in NC-IGF1RKD cells but no reduction of IR protein expression. Bar graphs show densitometry expressed as the mean fold change +/− SEM, t-test, ∗∗∗∗ p < 0.0001, n = 18 independent experiments. (B) Phosphorylation of AKT and p44/42MAPK in response to acute IGF1 stimulation at 10 ng and 100 ng/mL for 10 min was significantly reduced in NC-IGF1RKD podocytes. Data are expressed as the mean ± SEM, one-way ANOVA with Tukey’s multiple comparison test, ∗∗ p < 0.005, n = 3 independent experiments. (C) Western blot shows that IGF1R expression is reduced by ∼70% in wild-type podocytes exposed to 100 nM picropodophyllin for 24 h. Data are expressed as the mean ± SEM, t-test, ∗ p < 0.05, n = 3 independent experiments. No significant change in IR expression was observed. (D) Western blot shows the phosphorylation of AKT and p44/42MAPK in response to acute IGF1 stimulation at 10 ng and 100 ng/mL for 10 min in podocytes exposed to 100 nM picropodophyllin for 24 h. Data expressed as the mean ± SEM, ∗ p < 0.05, n = 3 independent experiments. (E) ∼50% of NC-IGF1RKD cells survive 7 days after gene excision. Treatment of wild-type podocytes with 100 nM picropodophyllin for 24 h has no effect on cell survival. Data are expressed as the mean ± SEM, t-test, ∗∗ p < 0.005, n = 3–4 independent experiments. See also Figures S5 and .

Article Snippet: For acute insulin and IGF1 stimulation, Conditionally immortalised wild-type mouse podocyte cell were serum starved for 4 h then 10 nM and 100 nM of insulin (Biotechne, Cat# 3435) or 10 ng/mL and 100 ng/mL IGF1 (Novus, Cat# NBP2-35081) was applied to the cells for 10 min.

Techniques: In Vitro, Activity Assay, Inhibition, Western Blot, Expressing, Comparison

Journal: iScience

Article Title: Contrasting consequences of podocyte insulin-like growth factor 1 receptor inhibition

doi: 10.1016/j.isci.2024.109749

Figure Lengend Snippet:

Article Snippet: For acute insulin and IGF1 stimulation, Conditionally immortalised wild-type mouse podocyte cell were serum starved for 4 h then 10 nM and 100 nM of insulin (Biotechne, Cat# 3435) or 10 ng/mL and 100 ng/mL IGF1 (Novus, Cat# NBP2-35081) was applied to the cells for 10 min.

Techniques: Recombinant, Protease Inhibitor, Enzyme-linked Immunosorbent Assay, Quantitation Assay, Staining, Western Blot, Cell Culture, Software, Plasmid Preparation

Journal: iScience

Article Title: Contrasting consequences of podocyte insulin-like growth factor 1 receptor inhibition

doi: 10.1016/j.isci.2024.109749

Figure Lengend Snippet: In vitro differential suppression of podocyte IGF1R activity reveals that partial inhibition is beneficial but near total loss is highly detrimental (A) Representative Western blot shows >90% reduction of IGF1R protein in NC-IGF1RKD cells but no reduction of IR protein expression. Bar graphs show densitometry expressed as the mean fold change +/− SEM, t-test, ∗∗∗∗ p < 0.0001, n = 18 independent experiments. (B) Phosphorylation of AKT and p44/42MAPK in response to acute IGF1 stimulation at 10 ng and 100 ng/mL for 10 min was significantly reduced in NC-IGF1RKD podocytes. Data are expressed as the mean ± SEM, one-way ANOVA with Tukey’s multiple comparison test, ∗∗ p < 0.005, n = 3 independent experiments. (C) Western blot shows that IGF1R expression is reduced by ∼70% in wild-type podocytes exposed to 100 nM picropodophyllin for 24 h. Data are expressed as the mean ± SEM, t-test, ∗ p < 0.05, n = 3 independent experiments. No significant change in IR expression was observed. (D) Western blot shows the phosphorylation of AKT and p44/42MAPK in response to acute IGF1 stimulation at 10 ng and 100 ng/mL for 10 min in podocytes exposed to 100 nM picropodophyllin for 24 h. Data expressed as the mean ± SEM, ∗ p < 0.05, n = 3 independent experiments. (E) ∼50% of NC-IGF1RKD cells survive 7 days after gene excision. Treatment of wild-type podocytes with 100 nM picropodophyllin for 24 h has no effect on cell survival. Data are expressed as the mean ± SEM, t-test, ∗∗ p < 0.005, n = 3–4 independent experiments. See also Figures S5 and .

Article Snippet: For acute insulin and IGF1 stimulation, Conditionally immortalised wild-type mouse podocyte cell were serum starved for 4 h then 10 nM and 100 nM of insulin (Biotechne, Cat# 3435) or 10 ng/mL and 100 ng/mL IGF1 (Novus, Cat# NBP2-35081) was applied to the cells for 10 min.

Techniques: In Vitro, Activity Assay, Inhibition, Western Blot, Expressing, Comparison

Journal: iScience

Article Title: Contrasting consequences of podocyte insulin-like growth factor 1 receptor inhibition

doi: 10.1016/j.isci.2024.109749

Figure Lengend Snippet:

Article Snippet: For acute insulin and IGF1 stimulation, Conditionally immortalised wild-type mouse podocyte cell were serum starved for 4 h then 10 nM and 100 nM of insulin (Biotechne, Cat# 3435) or 10 ng/mL and 100 ng/mL IGF1 (Novus, Cat# NBP2-35081) was applied to the cells for 10 min.

Techniques: Recombinant, Protease Inhibitor, Enzyme-linked Immunosorbent Assay, Quantitation Assay, Staining, RNAscope, Western Blot, Cell Culture, Software, Plasmid Preparation

Journal: iScience

Article Title: Contrasting consequences of podocyte insulin-like growth factor 1 receptor inhibition

doi: 10.1016/j.isci.2024.109749

Figure Lengend Snippet: In vitro differential suppression of podocyte IGF1R activity reveals that partial inhibition is beneficial but near total loss is highly detrimental (A) Representative Western blot shows >90% reduction of IGF1R protein in NC-IGF1RKD cells but no reduction of IR protein expression. Bar graphs show densitometry expressed as the mean fold change +/− SEM, t-test, ∗∗∗∗ p < 0.0001, n = 18 independent experiments. (B) Phosphorylation of AKT and p44/42MAPK in response to acute IGF1 stimulation at 10 ng and 100 ng/mL for 10 min was significantly reduced in NC-IGF1RKD podocytes. Data are expressed as the mean ± SEM, one-way ANOVA with Tukey’s multiple comparison test, ∗∗ p < 0.005, n = 3 independent experiments. (C) Western blot shows that IGF1R expression is reduced by ∼70% in wild-type podocytes exposed to 100 nM picropodophyllin for 24 h. Data are expressed as the mean ± SEM, t-test, ∗ p < 0.05, n = 3 independent experiments. No significant change in IR expression was observed. (D) Western blot shows the phosphorylation of AKT and p44/42MAPK in response to acute IGF1 stimulation at 10 ng and 100 ng/mL for 10 min in podocytes exposed to 100 nM picropodophyllin for 24 h. Data expressed as the mean ± SEM, ∗ p < 0.05, n = 3 independent experiments. (E) ∼50% of NC-IGF1RKD cells survive 7 days after gene excision. Treatment of wild-type podocytes with 100 nM picropodophyllin for 24 h has no effect on cell survival. Data are expressed as the mean ± SEM, t-test, ∗∗ p < 0.005, n = 3–4 independent experiments. See also Figures S5 and .

Article Snippet: Recombinant Mouse IGF1 protein , Novus , Cat# NBP2-35081.

Techniques: In Vitro, Activity Assay, Inhibition, Western Blot, Expressing, Comparison

Journal: iScience

Article Title: Contrasting consequences of podocyte insulin-like growth factor 1 receptor inhibition

doi: 10.1016/j.isci.2024.109749

Figure Lengend Snippet:

Article Snippet: Recombinant Mouse IGF1 protein , Novus , Cat# NBP2-35081.

Techniques: Recombinant, Protease Inhibitor, Enzyme-linked Immunosorbent Assay, Quantitation Assay, Staining, RNAscope, Western Blot, Cell Culture, Software, Plasmid Preparation

Journal: bioRxiv

Article Title: Experimental type 1 diabetes metabolically rejuvenates CD8 + T cells for improved control of tumor growth through an IGF1-IGF1R axis

doi: 10.1101/2024.04.04.588206

Figure Lengend Snippet: Expression of igf1, igf1R, igf2R and insulinR was analyzed by RT-PCR from ( A ) MACS sorted CD8 + TILs, ( B ) tumor cells and ( C ) MACS sorted CD8 + T cells isolated from TDLNs (n=6 mice/group). ( D) Flow cytometry gating and (E) percentage of CD8 + IGF1 + IGF1R + cells in tumors (n=4). CD8 + IGF1 + IGF1R + cells in (F) LNs and TDLNs, (G) spleen and (H) PBMCs (n=4 mice/group). (I) Correlation analysis between tumor area and the frequency of CD8 + IGF1 + IGF1R + T cells in tumors. (J) Colocalization of CD8 and IGF1 (arrow marked) in B16F10 tumor sections harvested from control vs. STZ induced T1D mice. (K) Colocalization of CD8 and IGF1R (arrow marked) in B16F10 tumor sections harvested from control vs. STZ induced T1D mice. Two-way ANOVA was performed to test for significance across groups. (*P ≤ 0.05, **P ≤ 0.01, ***P ≤ 0.001, ****P ≤ 0.0001)

Article Snippet: Rapamycin (Merck) (25 pM dissolved in DMSO) was used to block mTORC1 and mouse recombinant IGF1 (R&D Systems) (0.1 μg/ml) was used to induce IGF1R signaling. siIGF1R, rapamycin, and m-rIGF1 were administered alone and in all possible combinations in vitro to CD8 + T cells isolated from the TDLNs of diabetic and non-diabetic mice.

Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, Isolation, Flow Cytometry, Control

Journal: bioRxiv

Article Title: Experimental type 1 diabetes metabolically rejuvenates CD8 + T cells for improved control of tumor growth through an IGF1-IGF1R axis

doi: 10.1101/2024.04.04.588206

Figure Lengend Snippet: ( A) Western blot analysis of pAKT, AKT, mTOR and phosphor-mTORC1 (p-mTORC1) in MACS sorted CD8 + TILs (n=3). ( B) Search tool for the retrieval of interacting genes/proteins (STRING) analysis was used to study signaling interactions between IGF1, IGF1R, InsulinR and mTOR. ( C) Schematic overview of in vitro experiments (n=3) to study cellular signaling in MACS sorted CD8 + T cells isolated from TDLN. rIGF1 was used to stimulate the IGF1-IGF1R-mTOR signaling axis, siIGF1R and rapamycin were used to block IGF1R expression and mTORC1, respectively. ( D) Expression of igf1, igf1R, glut1, ldha, gzmB and ifnγ following in vitro treatment of rIGF1, siIGF1R and rapamycin, either alone or in combination. (E) Percent CD8 + CD69 + cells in Cont. CD8 + T cells vs. STZ CD8 + T cells (n=3). (F) Percent CD8 + IFNψ + cells amongst Cont. CD8 + vs. STZ CD8 groups (n=3). (G) Flow cytometry gating on CD8 + , CD69 + and IGF1R + cells in T1D patient PBMC. (H) Percent CD8 + CD69 + and (I) percent CD8 + CD69 + cell population in T1D patient and non-diabetic PBMC samples after in vitro treatment with tumor lysate (TL), rapamycin and picropodophyllotoxin (PPT) alone or in combination. Two-way ANOVA followed by Tukey’s multiple comparison test was performed to test significance of intergroup differences. (*P ≤ 0.05, **P ≤ 0.01, ***P ≤ 0.001, ****P ≤ 0.0001)

Article Snippet: Rapamycin (Merck) (25 pM dissolved in DMSO) was used to block mTORC1 and mouse recombinant IGF1 (R&D Systems) (0.1 μg/ml) was used to induce IGF1R signaling. siIGF1R, rapamycin, and m-rIGF1 were administered alone and in all possible combinations in vitro to CD8 + T cells isolated from the TDLNs of diabetic and non-diabetic mice.

Techniques: Western Blot, In Vitro, Isolation, Blocking Assay, Expressing, Flow Cytometry, Comparison