igf1  (Novus Biologicals)

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