recombinant murine igf1  (PeproTech)

Journal: Journal of Cellular and Molecular Medicine

Article Title: EGFR marks a subpopulation of dermal mesenchymal cells highly expressing IGF1 which enhances hair follicle regeneration

doi: 10.1111/jcmm.17766

Figure Lengend Snippet: Analysis of scRNA‐seq data to identify dermal mesenchymal cell population that co‐expresses Egfr and Igf1 in neonatal murine skin. (A) UMAP visualisation of all cell populations in 2‐day‐old neonatal mouse skin. Each point represents a single cell, colour‐coded based on cell types. (B) Subset and clustering of dermal cells. (C) Visualisation of dermal and epithelial marker gene expression across dermal subtypes shown in the dot plot. (D) Vlnplot showing Egfr and Igf1 expression in dermal subclusters. (E) Heatmap of top 50 differently expressed genes in EGFR − cells, EGFR + cells and dermal papilla (DP) cells. (F) The GO terms of the BP, CC, and MF categories enrichment of the upregulated genes in EGFR + cells. Within Cartesian coordinates (x,y), the closer the different circles are, the more related the GO terms are. The size of the circle is proportional to the number of genes within the GO terms. The colour of the circle indicates the significance enrichment. BP, biological process; CC, cellular component; GO, Gene Ontology; MF, molecular function.

Article Snippet: From post‐injury Day 7 to Day 14, each mouse received a daily injection of 50 μL PBS or murine IGF1 (MCE, HY‐P7070) solution (dissolved in PBS at 1 ng/μl) in the two wounds, respectively (beneath the scab into the wound bed tissue).

Techniques: Marker, Expressing

Journal: Journal of Cellular and Molecular Medicine

Article Title: EGFR marks a subpopulation of dermal mesenchymal cells highly expressing IGF1 which enhances hair follicle regeneration

doi: 10.1111/jcmm.17766

Figure Lengend Snippet: Isolation of EGFR expressing mesenchymal cells present in neonatal murine dermis. (A) Immunofluorescence staining for EGFR in tetracycline‐induced neonatal skin from Krt14‐rtTA‐H2BGFP. Scale bar: 50 μm. White arrows point to EGFR positive non‐epithelial cells. (B) Schematic illustration of our strategy for analysing and sorting of EGFR‐expressing mesenchymal cells in neonatal dermis. (C) Fluorescence‐activated cell sorting (FACS) of EGFR+ cells in compo‐site‐FITC negative cells. APC, allophycocyanin; GFP, green fluorescent protein. (D) Real‐Time PCR analysis of mesenchymal marker (Col1a1) expression in freshly isolated mouse dermal subpopulations (Data represents mean ± SEM, n = 3). (E) Immunofluorescence staining of EGFR, vimentin (VIM), DAPI. Scale bar, 100 μm. (F) Real‐time PCR analysis of Igf1 expression in the freshly isolated mouse dermal subpopulations. In Real‐time PCR experiments (D and F), we used triple wells for each sample. Samples from two independent experiments were examined and showed similar results. Results of one analysis are shown. ** p < 0.01, *** p < 0.001; EGFR‐ and EGFR+, represent EGFR negative and positive cells; Epi, epidermal cells; ns, means no significant difference.

Article Snippet: From post‐injury Day 7 to Day 14, each mouse received a daily injection of 50 μL PBS or murine IGF1 (MCE, HY‐P7070) solution (dissolved in PBS at 1 ng/μl) in the two wounds, respectively (beneath the scab into the wound bed tissue).

Techniques: Isolation, Expressing, Immunofluorescence, Staining, Fluorescence, FACS, Real-time Polymerase Chain Reaction, Marker

Journal: Journal of Cellular and Molecular Medicine

Article Title: EGFR marks a subpopulation of dermal mesenchymal cells highly expressing IGF1 which enhances hair follicle regeneration

doi: 10.1111/jcmm.17766

Figure Lengend Snippet: EGFR and IGF1 expression in adult murine dermis. (A) Immunofluorescence staining for EGFR and DAPI in skin tissues of 7‐week‐old PDGFRa cre ERT;mTmG mice . Scale bar, 100 μm. (B) Immunofluorescence staining of EGFR in tetracycline‐induced Krt14‐rtTA‐H2BGFP adult mouse skin. White arrows point to EGFR positive non‐epithelial cells. Scale bars, 100 μm (C) Immunofluorescence staining of IGF1 in Day 7 wounds. Scale bars, 200 μm. (D) Co‐localisation of IGF1 and EGFR in adult wound dermis. White arrows point to EGFR positive cells. Scale bars, 100 μm.

Article Snippet: From post‐injury Day 7 to Day 14, each mouse received a daily injection of 50 μL PBS or murine IGF1 (MCE, HY‐P7070) solution (dissolved in PBS at 1 ng/μl) in the two wounds, respectively (beneath the scab into the wound bed tissue).

Techniques: Expressing, Immunofluorescence, Staining

Journal: Journal of Cellular and Molecular Medicine

Article Title: EGFR marks a subpopulation of dermal mesenchymal cells highly expressing IGF1 which enhances hair follicle regeneration

doi: 10.1111/jcmm.17766

Figure Lengend Snippet: Effects of IGF1 on wound‐induced hair follicle neogenesis. (A) Schematic representation of strategy for exogenous IGF1 injection during wound healing process. Two full thickness wound (diameter = 2 mm) were made on neonatal mouse dorsal skin. From post‐wounding Day 7 to Day 14, each mouse received a daily injection of 50 μL PBS or murine IGF1 solution (dissolved in PBS at 1 ng/μl) beneath the scab into the wound bed tissue in the two dorsal wounds, respectively. (B) AP staining on wound tissue from control group (PBS) and IGF1 injection group collected at post wound Day 15. The wound bed lies between red dash lines in each picture. AP was stained blue. Red arrows point to AP positive area including neogenic hair follicles. Scale bar, 200 μm. (C) Visualisation of hair follicles in healed wounds collected at Day 21. (D) Statistics of hair follicles in both groups collected at Day 21. Data represents the mean ± SEM, n = 4, * p < 0.05.

Article Snippet: From post‐injury Day 7 to Day 14, each mouse received a daily injection of 50 μL PBS or murine IGF1 (MCE, HY‐P7070) solution (dissolved in PBS at 1 ng/μl) in the two wounds, respectively (beneath the scab into the wound bed tissue).

Techniques: Injection, Staining, Control

Journal: Science Advances

Article Title: Hedgehog-interacting protein orchestrates alveologenesis and protects against bronchopulmonary dysplasia and emphysema

doi: 10.1126/sciadv.adu2958

Figure Lengend Snippet: ( A ) IF analysis of SMA in the alveoli of Hhip -deleted (HHIP CKO) and control mice at P14. ( B ) Number of myofibroblasts per unit alveolar area of Hhip -deleted and control mice. ( C and D ) UMAP showing cell clusters in the lung fibroblasts of Hhip -deleted and control mice at P14. ( E ) Violin plots showing the expression of Pdgfra , Acta2 , Hhip , and Cdh4 in alveolar myofibroblasts (ALMF), ductal myofibroblasts (DMF), peribronchial fibroblasts (Perib), adventitial fibroblasts (Adv), and alveolar fibroblasts (Alv). ( F ) Expression of Acta2 , Myh11 , Tagln , and Igf1 in ALMFs and DMFs of Hhip -deleted and control mice. ( G ) qPCR analysis of Igf1 expression in the lung stromal cells isolated from Hhip -deleted and control mice. ( H ) Analysis of Igf1 (RNA in situ ) and SMA expression in the alveoli. ( I ) Number of SMA + Igf1 + cells per unit alveolar area of Hhip -deleted and control mice at P14. ( J ) Percentage of SMA + Igf1 + cells in total Igf1 + cells. ( K ) Top 10 activated pathways in Hhip -deleted, relative to control myofibroblasts, analyzed with IPA. ( L ) qPCR analysis of Gli1 , Igf1 , and Acta2 expression in the lung stromal cells treated with PBS, SHH, and SHH plus HHIP. ( M ) qPCR analysis of Acta2 expression in SHH-stimulated lung stromal cells treated with vehicle or IGF1R inhibitor. ( N ) IF analysis of SMA expression in the alveoli of Hhip -deleted mice administered with vehicle or IGF1R inhibitor. ( O ) Number of myofibroblasts per unit alveolar area of Hhip -deleted mice administered with vehicle or IGF1R inhibitor. All in vitro experiments have been repeated at least one time with consistent results for validation. Each data point represents one mouse [(B), (G), (I), (J), and (O)] of an individual experiment. Data are expressed as mean ± SD. * P < 0.05, ** P < 0.005, *** P < 0.0005, and **** P < 0.0001.

Article Snippet: For IGF-1 treatment, the confluent fibroblasts were cultured in 0.5% FBS with 1% antibiotic-antimycotic, and recombinant murine IGF1 (catalog no. 250-19; PeproTech) was added at 50 ng/ml.

Techniques: Control, Expressing, Isolation, In Situ, In Vitro, Biomarker Discovery

Journal: Science Advances

Article Title: Hedgehog-interacting protein orchestrates alveologenesis and protects against bronchopulmonary dysplasia and emphysema

doi: 10.1126/sciadv.adu2958

Figure Lengend Snippet: ( A ) Violin plots showing the expression of Cdkn1a in ALMFs and DMFs of Hhip -deleted and control mice. ( B ) Senescence β-galactosidase staining of Hhip -deleted and control lungs. ( C ) IF analysis of SMA, p21, and CDH4 in Hhip -deleted and control lungs. Arrow: p21 + DMFs; arrowhead: p21 + ALMFs. ( D and E ) Number of p21 + ALMFs (D) and DMFs (E) per unit alveolar area of Hhip -deleted and control mice at P14. ( F ) IF analysis of SPC and p21 in Hhip -deleted and control lungs. Arrow: p21 + SPC + cells. ( G ) Percentage of p21 + cells in AT2s. ( H ) Activation of senescence pathways in the AT2s of Hhip -deleted mice, relative to control AT2s, and analyzed with IPA. ( I ) Top 5 upstream regulators in the AT2s of Hhip -deleted mice, relative to control AT2s, analyzed with IPA. ( J ) AT2 organoids cocultured with lung stromal cells ( R26R SmoM2/+ ) pre-infected with adenovirus-empty and adenovirus-Cre, treated with anti-IGF1 antibody and IgG. ( K and L ) Quantification of colony-forming efficiency (CFE) and organoid size. ( M ) IF analysis of SPC and p21 in AT2 organoids. Arrow: p21 + SPC + cells. ( N ) Percentage of p21 + cells in AT2s in the organoid assay. All in vitro experiments have been repeated at least one time with consistent results for validation. Each data point represents one mouse [(D), (E), and (G)] of an individual experiment. Data are expressed as Mean ± SD. * P < 0.05, ** P < 0.005, *** P < 0.0005, and **** P < 0.0001.

Article Snippet: For IGF-1 treatment, the confluent fibroblasts were cultured in 0.5% FBS with 1% antibiotic-antimycotic, and recombinant murine IGF1 (catalog no. 250-19; PeproTech) was added at 50 ng/ml.

Techniques: Expressing, Control, Staining, Activation Assay, Infection, In Vitro, Biomarker Discovery

Journal: Science Advances

Article Title: Hedgehog-interacting protein orchestrates alveologenesis and protects against bronchopulmonary dysplasia and emphysema

doi: 10.1126/sciadv.adu2958

Figure Lengend Snippet: ( A ) snRNA-seq analysis of GLI1 , PATCH1 , IGF1 , and ACTA2 expression in ALMFs and DMFs of human BPD. ( B ) H&E images of the lungs of hyperoxia-treated and control mice. ( C ) Quantification of MLI of hyperoxia-treated and control lungs. ( D ) qPCR analysis of Hhip , Gli1 , and Igf1 expression in the lung stromal cells isolated from hyperoxia-treated and control mice. ( E ) IF analysis of SMA in the lungs of hyperoxia-treated and control mice. ( F ) Histology quantification of the number of myofibroblasts of hyperoxia-treated and control mice. ( G ) qPCR analysis of Acta2 expression in the lung stromal cells isolated from hyperoxia-treated and control mice. ( H ) qPCR analysis of Cdkn1a expression in the lung epithelial cells isolated from hyperoxia-treated and control mice. ( I ) IF analysis of p21 and SPC in the lungs of hyperoxia-treated and control mice. Arrow: p21 + SPC + cells. ( J and K ) Quantification of the percentage of p21 + AT2s and number of AT2s in hyperoxia-treated and control mice. Each data point represents one mouse [(C), (D), (F) to (H), (J), and (K)] of an individual experiment. Data are expressed as mean ± SD. * P < 0.05, ** P < 0.005, and **** P < 0.0001.

Article Snippet: For IGF-1 treatment, the confluent fibroblasts were cultured in 0.5% FBS with 1% antibiotic-antimycotic, and recombinant murine IGF1 (catalog no. 250-19; PeproTech) was added at 50 ng/ml.

Techniques: Expressing, Control, Isolation

Journal: Science Advances

Article Title: Hedgehog-interacting protein orchestrates alveologenesis and protects against bronchopulmonary dysplasia and emphysema

doi: 10.1126/sciadv.adu2958

Figure Lengend Snippet: ( A ) Design strategy for HHIP-Fc recombinant protein and its analysis by Western blotting. ( B ) qPCR analysis of Gli1 expression in the lung stromal cells isolated from neonatal mice 3 and 5 days after one dose of HHIP or HHIP-Fc treatment. ( C ) H&E images of hyperoxia-exposed lungs, treated with HHIP-Fc or Fc fragment control. ( D ) MLI quantification of hyperoxia-exposed lungs, treated with HHIP-Fc or Fc. ( E and F ) IF analysis and quantification of myofibroblasts in hyperoxia-exposed lungs, treated with HHIP-Fc or Fc. ( G to I ) IF analysis and quantification of AT2 number and p21 + AT2 percentage in hyperoxia-exposed lungs, treated with HHIP-Fc or Fc. Arrow: p21 + SPC + cells. ( J ) qPCR analysis of Igf1 expression in the lung stromal cells isolated from hyperoxia-exposed mice, treated with HHIP-Fc or Fc. Each data point represents one mouse [(B), (D), (F), and (H) to (J)] of an individual experiment. Data are expressed as mean ± SD. * P < 0.05, ** P < 0.005, *** P < 0.0005, and **** P < 0.0001.

Article Snippet: For IGF-1 treatment, the confluent fibroblasts were cultured in 0.5% FBS with 1% antibiotic-antimycotic, and recombinant murine IGF1 (catalog no. 250-19; PeproTech) was added at 50 ng/ml.

Techniques: Recombinant, Western Blot, Expressing, Isolation, Control

Journal: Glia

Article Title: Intranasal mesenchymal stem cell therapy to boost myelination after encephalopathy of prematurity

doi: 10.1002/glia.23919

Figure Lengend Snippet: Overview of primer sequences used in validation qPCR

Article Snippet: To the OPC cultures, the following selected factors were added: Noggin (murine recombinant, Peprotech 250‐38), IL10 (rat recombinant, Peprotech 400‐19), IL11 (murine recombinant, Peprotech 220‐11), LIF (murine recombinant, Peprotech 250‐02), CXCL12 (murine recombinant, Peprotech 250‐20a), GCSF (murine recombinant, Peprotech 250‐05), IGF1 (murine recombinant, Peprotech 250‐19) and EGF (murine recombinant, Peprotech 250‐09).

Techniques:

Journal: Glia

Article Title: Intranasal mesenchymal stem cell therapy to boost myelination after encephalopathy of prematurity

doi: 10.1002/glia.23919

Figure Lengend Snippet: The MSCs secretome boosts OL maturation and attenuates microglia activation in vitro. (a) MCM + LPS causes a reduction in MBP + area (dashed line represents MBP + area in MCM‐LPS control condition), MSC treatment with 4 × 10 4 or 8 × 10 4 MSCs in a noncontact co‐culture significantly improves OL maturation ( n = 3 independent experiments, 3–4 observations per experiment normalized for the positive control, for example, cells exposed to MCM + LPS). (b) Representative fluorescent images (×10) of primary cultured oligodendrocytes stained for oligodendrocyte marker Olig2 (red) and myelin component MBP (green). Cells were exposed to MCM‐LPS (MCM−) or MCM + LPS (MCM+) and 4 × 10 4 MSCs (MSC+) in a noncontact gel‐insert. Scale bars: 100 μm. (c) Exposure to 10 ng/ml TNFα leads to a reduction in MBP production (dashed line represents MBP + area in medium without TNFα), MSC treatment with 4x10 4 MSCs in a noncontact co‐culture significantly boosts OL maturation ( n = 2 independent experiments, 3–4 observations per experiment, normalized for the positive control, for example, cells exposed to TNFα). (d) Representative fluorescent images (×10) of primary cultured oligodendrocytes stained for oligodendrocyte marker Olig2 (red) and myelin component MBP (green). Cells were exposed to medium with (+) or without (−) 10 ng/ml TNFα and 4 × 10 4 MSCs (MSC+) in noncontact gel‐insert. Scale bars: 100 μm. (e). Treatment with 4 × 10 4 MSCs in a noncontact gel‐insert attenuates microglial TNFα production ( n = 2 independent experiments, two observations per experiment, normalized for the positive control, for example, cells exposed to LPS). (f,g) Addition of IGF1, EGF, Noggin, GCSF, LIF, and IL11 but not IL10 and CXCL12 significantly improves MBP + area by primary cultured oligodendrocytes following MCM + LPS exposure (dashed line represents MBP + area in MCM‐LPS control condition) ( n = 2 independent experiments, 3–4 observations per experiment normalized for the positive control, for example, cells exposed to MCM + LPS). (h,i) Addition of IGF1, EGF, LIF, IL11, and CXCL12 but not Noggin, G‐CSF, and IL10 boosts OL maturation after TNFα‐induced OL maturational arrest (dashed line represents MBP + area in medium without TNFα) ( n = 2 independent experiments, 3–4 observations per experiment, normalized for the positive control, for example, cells exposed to TNFα). ## p < .01; ### p < .001; #### p < .0001 MCM+ or TNF+ condition (black bars) versus MCM‐ or TNFα‐ control (dashed line), respectively; * p < .05; ** p < .01; *** p < .001; **** p < .0001 factor‐exposed MCM+ or TNF+ condition (gray bars) versus MCM+ or TNF+ control condition (black bars) [Color figure can be viewed at wileyonlinelibrary.com ]

Article Snippet: To the OPC cultures, the following selected factors were added: Noggin (murine recombinant, Peprotech 250‐38), IL10 (rat recombinant, Peprotech 400‐19), IL11 (murine recombinant, Peprotech 220‐11), LIF (murine recombinant, Peprotech 250‐02), CXCL12 (murine recombinant, Peprotech 250‐20a), GCSF (murine recombinant, Peprotech 250‐05), IGF1 (murine recombinant, Peprotech 250‐19) and EGF (murine recombinant, Peprotech 250‐09).

Techniques: Activation Assay, In Vitro, Co-Culture Assay, Positive Control, Cell Culture, Staining, Marker

Journal: Glia

Article Title: Intranasal mesenchymal stem cell therapy to boost myelination after encephalopathy of prematurity

doi: 10.1002/glia.23919

Figure Lengend Snippet: Trophic gene expression changes in dWMI animals after MSC treatment versus vehicle treatment

Article Snippet: To the OPC cultures, the following selected factors were added: Noggin (murine recombinant, Peprotech 250‐38), IL10 (rat recombinant, Peprotech 400‐19), IL11 (murine recombinant, Peprotech 220‐11), LIF (murine recombinant, Peprotech 250‐02), CXCL12 (murine recombinant, Peprotech 250‐20a), GCSF (murine recombinant, Peprotech 250‐05), IGF1 (murine recombinant, Peprotech 250‐19) and EGF (murine recombinant, Peprotech 250‐09).

Techniques: Expressing