igf1 r beta  (Cell Signaling Technology Inc)

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: 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