Journal: Nature Communications

Article Title: Sensory nerve niche regulates mesenchymal stem cell homeostasis via FGF/mTOR/autophagy axis

doi: 10.1038/s41467-023-35977-4

Figure Lengend Snippet: a Expression of FGF1 and β-gal in one-month-old Gli1-LacZ mice. b Relative contributions of ligand-receptor pairs to the overall communication network of FGF signaling pathway; the Fgf1-Fgfr1 pair is the major contributor. c Expression of FGFR1 and β-gal in one-month-old Gli1-LacZ mice. d Immunoprecipitation assay demonstrating the interaction between FGF1 and FGFR1 in the proximal mesenchyme of the incisor. e Expression of Fgfr1 and tdTomato in MSC cultures from Gli1 CreER ;tdTomato mice. f – j Abnormal dentin formation in Gli1 CreER ;Fgfr1 fl/fl mice three months after tamoxifen induction. f CT imaging of the incisors of Fgfr1 fl/fl and Gli1 CreER ;Fgfr1 fl/fl mice. White arrow points to the dental pulp cavity; the white arrowhead points to narrowed pulp cavity. g Histological analysis of Fgfr1 fl/fl and Gli1 CreER ;Fgfr1 fl/fl mice. Yellow arrow points to pre-odontoblast; yellow arrowhead points to abnormal pre-odontoblast; asterisk points to abnormal dentin formation. h Expression of Dspp in control and Gli1 CreER ;Fgfr1 fl/fl mice. Yellow arrow points to the distance between the bending point of the cervical loop and the initiation of odontoblast differentiation. i Quantification of dental pulp cavity percentage in control and mutant mice. P = 0.0008. j Quantification of the distance of Dspp + cells to cervical loop in control and mutant mice. P < 0.0001. Schematic at the bottom indicates the induction protocol. mpt month post-tamoxifen injection. k The growth rate of the incisor was detected with notch movement observed at day (D)3, D6, and D14 in control and Gli1 CreER ;Fgfr1 fl/fl mice. White arrow points to the notch location; the schematic at the bottom indicates the induction protocol. mpt month post-tamoxifen injection. l Quantification of notch movement in control and mutant mice. P = 0.0123. For I , j and l , n = 3 and each data point represent one animal, with unpaired Student’s t -test performed. All data are expressed as the mean ± SD. Source data are provided as a Source Data file. *** P < 0.001, **** P < 0.0001. Each experiment was repeated independently three times. White dotted line outlines the cervical loop. Cre-: Fgfr1 fl/fl ; Cre+: Gli1 CreER ;Fgfr1 fl/fl . Scale bars, f 1 mm; k 2 mm; e 50 μm; others, 100 μm.

Article Snippet: Membranes were blocked with 5% non-fat dry milk for 1 h, then incubated with primary antibodies: anti-FGFR1 (Cell signaling technology 9740, 1:1000), anti-FGF1 (Abcam ab207321, 1:1000), anti-LC3 (1:1000, Abcam, ab48394), anti-p-ULK1 (Cell Signaling technology 14202, 1:1000), anti-p-mTOR (Cell Signaling technology 5536, 1:1000), anti-mTOR (Cell Signaling technology 2972, 1:1000), anti-p-p70S6K (Cell Signaling technology 9204, 1:1000), anti-p70S6K (Cell Signaling technology 2708, 1:1000), anti-p-S6 (Cell Signaling technology 4858, 1:1000), anti-S6 (Cell Signaling technology 2217, 1:1000), anti-p-Akt (Cell Signaling technology 4060, 1:1000), anti-Akt (Cell Signaling technology 9272, 1:1000), anti-p-p38 (Cell Signaling technology 4511, 1:1000), anti-p38 (Cell Signaling technology 8690, 1:1000), anti-p-ERK (Cell Signaling technology 4370, 1:1000), anti-ERK (Cell Signaling technology 4695, 1:1000), anti-p-JNK (Cell Signaling technology 9255, 1:1000), anti-JNK (Cell Signaling technology 9252, 1:1000), and anti-β-actin (Abcam ab20272, 1:1000) at 4 °C overnight.

Techniques: Expressing, Immunoprecipitation, Imaging, Mutagenesis, Injection

Journal: Nature Communications

Article Title: Sensory nerve niche regulates mesenchymal stem cell homeostasis via FGF/mTOR/autophagy axis

doi: 10.1038/s41467-023-35977-4

Figure Lengend Snippet: a–d The number of MSCs decreased in Gli1 CreER ;Fgfr1 fl/fl mice one week after tamoxifen induction. Gli1 + cells stained with β-gal in incisors of control and Gli1 CreER ;Fgfr1 fl/fl mice. e , Quantification of the percentage of Gli1 + cells in control and mutant mice. P = 0.0007. f–i TACs detected with Ki67 staining in control and Gli1 CreER ;Fgfr1 fl/fl mice. j Quantification of Ki67 + TACs in control and mutant mice. P = 0.0008. k–n The expression of Dspp and EdU in control and Fgfr1 fl/fl mutant mice. The length of overlap between Dspp + odontoblasts and EdU + cells reflects the number of TACs undergoing odontoblastic differentiation. White arrowhead points to overlap between Dspp + odontoblasts and EdU + cells. o Quantification of overlap between Dspp + odontoblasts and EdU + cells in control and mutant mice. P = 0.0011. p and q The migration of Gli1 + cells’ progeny indicated with tdTomato in Gli1 CreER ;tdTomato and Gli1 CreER ;Fgfr1 fl/fl ; tdTomato mice. White arrows point to tdTomato + cell migration. Yellow dot line with arrowheads point to migration distance. r Quantification of the percentage of tdTomato length in control and mutant mice. P = 0.0005. For e, o and r , n = 3 and each data point represents one animal, with unpaired Student’s t -test performed. All data are expressed as the mean ± SD. Source data are provided as a Source Data file. ** P < 0.01, *** P < 0.001. Each experiment was repeated independently three times. White dotted line outlines the cervical loop. Scale bars, p and q 400 μm; others, 100 μm.

Article Snippet: Membranes were blocked with 5% non-fat dry milk for 1 h, then incubated with primary antibodies: anti-FGFR1 (Cell signaling technology 9740, 1:1000), anti-FGF1 (Abcam ab207321, 1:1000), anti-LC3 (1:1000, Abcam, ab48394), anti-p-ULK1 (Cell Signaling technology 14202, 1:1000), anti-p-mTOR (Cell Signaling technology 5536, 1:1000), anti-mTOR (Cell Signaling technology 2972, 1:1000), anti-p-p70S6K (Cell Signaling technology 9204, 1:1000), anti-p70S6K (Cell Signaling technology 2708, 1:1000), anti-p-S6 (Cell Signaling technology 4858, 1:1000), anti-S6 (Cell Signaling technology 2217, 1:1000), anti-p-Akt (Cell Signaling technology 4060, 1:1000), anti-Akt (Cell Signaling technology 9272, 1:1000), anti-p-p38 (Cell Signaling technology 4511, 1:1000), anti-p38 (Cell Signaling technology 8690, 1:1000), anti-p-ERK (Cell Signaling technology 4370, 1:1000), anti-ERK (Cell Signaling technology 4695, 1:1000), anti-p-JNK (Cell Signaling technology 9255, 1:1000), anti-JNK (Cell Signaling technology 9252, 1:1000), and anti-β-actin (Abcam ab20272, 1:1000) at 4 °C overnight.

Techniques: Staining, Mutagenesis, Expressing, Migration

Journal: Nature Communications

Article Title: Sensory nerve niche regulates mesenchymal stem cell homeostasis via FGF/mTOR/autophagy axis

doi: 10.1038/s41467-023-35977-4

Figure Lengend Snippet: a–i mTOR-dependent autophagy is downregulated in Gli1 CreER ;Fgfr1 fl/fl mice. a Hierarchical clustering of control and Gli1 CreER ;Fgfr1 fl/fl mice. b Volcano plot showing 2019 upregulated genes and 1447 downregulated genes. c Significant signaling pathways analyzed with Ingenuity Pathway Analysis (IPA). d Expression of FGFR1, LC3 and p-ULK1 in mesenchyme of incisors from control and Gli1 CreER ;Fgfr1 fl/fl mice. e–h Expression of autophagy substrate P62 in control and Gli1 CreER ;Fgfr1 fl/fl mice. i Expression of p-mTOR and its downstream effectors p-P70S6K and pS6 in control and mutant mice. j–p FGF/p-JNK signaling regulates mTOR-dependent autophagy activation in MSCs. j Western blot of p-AKT, AKT, p-P38, P38, p-ERK, ERK, p-JNK, and JNK in proximal mesenchyme from control and mutant mice. k and l Expression of p-JNK and β-gal in the proximal mesenchyme of incisors from Gli1-LacZ mice. m–p Expression of p-JNK in control and Gli1 CreER ;Fgfr1 fl/fl mice. q Relative protein level in ( d ). FGFR1/β-Actin: P = 0.0001; LC3II/LC3I: P = 0.0006; p-ULK1/β-Actin: P < 0.0001. r Relative protein level in ( i ). p-mTOR/mTOR: P = 0.0056; p-P70S6K/P70S6K: P = 0.0016; pS6/S6: P = 0.0001. s Relative protein level in ( j ). p-Akt/Akt: P = 0.3485; p-P38/P38: P = 0.9988; p-ERK/ERK: P = 0.0025; p-JNK/JNK: P < 0.0001. t Relative fluorescent intensity of P62 and p-JNK. P62: P < 0.0001; p-JNK: P = 0.0019. For q–t , n = 3 and each data point represent one biological replicate, with unpaired Student’s t -test performed. All data are expressed as the mean ± SD. Source data are provided as a Source Data file. ** P < 0.01, *** P < 0.001, **** P < 0.0001. Each experiment was repeated independently three times. White dotted line outlines the cervical loop. Scale bars, 100 μm.

Article Snippet: Membranes were blocked with 5% non-fat dry milk for 1 h, then incubated with primary antibodies: anti-FGFR1 (Cell signaling technology 9740, 1:1000), anti-FGF1 (Abcam ab207321, 1:1000), anti-LC3 (1:1000, Abcam, ab48394), anti-p-ULK1 (Cell Signaling technology 14202, 1:1000), anti-p-mTOR (Cell Signaling technology 5536, 1:1000), anti-mTOR (Cell Signaling technology 2972, 1:1000), anti-p-p70S6K (Cell Signaling technology 9204, 1:1000), anti-p70S6K (Cell Signaling technology 2708, 1:1000), anti-p-S6 (Cell Signaling technology 4858, 1:1000), anti-S6 (Cell Signaling technology 2217, 1:1000), anti-p-Akt (Cell Signaling technology 4060, 1:1000), anti-Akt (Cell Signaling technology 9272, 1:1000), anti-p-p38 (Cell Signaling technology 4511, 1:1000), anti-p38 (Cell Signaling technology 8690, 1:1000), anti-p-ERK (Cell Signaling technology 4370, 1:1000), anti-ERK (Cell Signaling technology 4695, 1:1000), anti-p-JNK (Cell Signaling technology 9255, 1:1000), anti-JNK (Cell Signaling technology 9252, 1:1000), and anti-β-actin (Abcam ab20272, 1:1000) at 4 °C overnight.

Techniques: Expressing, Mutagenesis, Activation Assay, Western Blot

Journal: Nature Communications

Article Title: Sensory nerve niche regulates mesenchymal stem cell homeostasis via FGF/mTOR/autophagy axis

doi: 10.1038/s41467-023-35977-4

Figure Lengend Snippet: a–e Abnormal dentin deposition seen in Gli1 CreER ;Fgfr1 fl/fl mice can be rescued by rapamycin treatment for 3 months. a CT scanning of control, Gli1 CreER ;Fgfr1 fl/fl mice treated with vehicle ( Gli1 CreER ;Fgfr1 fl/fl + Veh) and Gli1 CreER ;Fgfr1 fl/fl mice treated with rapamycin ( Gli1 CreER ;Fgfr1 fl/fl + Rap). White arrow points to the dental pulp cavity; white arrowhead points to narrowed pulp cavity. b Histological analysis of these three groups. Yellow arrow points to normal pre-odontoblast; yellow arrowhead points to abnormal pre-odontoblast; asterisk points to abnormal dentin formation. c Expression of Dspp in these three groups. Yellow arrow points to the distance between the bending point of the cervical loop and the initiation of odontoblast differentiation. d Quantification of dental pulp cavity percentage in these three groups. Control vs Gli1 CreER ;Fgfr1 fl/fl + Veh: P = 0.0001; Gli1 CreER ;Fgfr1 fl/fl + Veh vs. Gli1 CreER ;Fgfr1 fl/fl + Rap: P = 0.0006. e Quantification of the distance of Dspp + cells to cervical loop. Control versus Gli1 CreER ;Fgfr1 fl/fl + Veh: P < 0.0001; Gli1 CreER ;Fgfr1 fl/fl + Veh versus Gli1 CreER ;Fgfr1 fl/fl + Rap: P < 0.0001. f Re-activation of autophagy benefits the retention of MSCs. Gli1 + cells labeled with β-gal in control, Gli1 CreER ;Fgfr1 fl/fl mice treated with vehicle or rapamycin. g Quantification of the percentage of Gli1 + cells. Control versus Gli1 CreER ;Fgfr1 fl/fl + Veh: P = 0.0002; Gli1 CreER ;Fgfr1 fl/fl + Veh versus Gli1 CreER ;Fgfr1 fl/fl + Rap: P = 0.0002. h TACs detected with Ki67 staining in control, Gli1 CreER ;Fgfr1 fl/fl mice treated with vehicle or rapamycin. i Quantification of Ki67 + TAC cells. Control versus Gli1 CreER ;Fgfr1 fl/fl + Veh: P < 0.0001; Gli1 CreER ;Fgfr1 fl/fl + Veh versus Gli1 CreER ;Fgfr1 fl/fl + Rap: P = 0.0021. For d , e , g , and I , n = 3 biologically independent samples, each data point represents one animal, with unpaired one-way ANOVA analysis. All data are expressed as the mean ± SD. Source data are provided as a Source Data file. ** P < 0.01, *** P < 0.001, **** P < 0.0001. Each experiment was repeated independently three times. White dotted line outlines the cervical loop. Scale bars, 100 μm.

Article Snippet: Membranes were blocked with 5% non-fat dry milk for 1 h, then incubated with primary antibodies: anti-FGFR1 (Cell signaling technology 9740, 1:1000), anti-FGF1 (Abcam ab207321, 1:1000), anti-LC3 (1:1000, Abcam, ab48394), anti-p-ULK1 (Cell Signaling technology 14202, 1:1000), anti-p-mTOR (Cell Signaling technology 5536, 1:1000), anti-mTOR (Cell Signaling technology 2972, 1:1000), anti-p-p70S6K (Cell Signaling technology 9204, 1:1000), anti-p70S6K (Cell Signaling technology 2708, 1:1000), anti-p-S6 (Cell Signaling technology 4858, 1:1000), anti-S6 (Cell Signaling technology 2217, 1:1000), anti-p-Akt (Cell Signaling technology 4060, 1:1000), anti-Akt (Cell Signaling technology 9272, 1:1000), anti-p-p38 (Cell Signaling technology 4511, 1:1000), anti-p38 (Cell Signaling technology 8690, 1:1000), anti-p-ERK (Cell Signaling technology 4370, 1:1000), anti-ERK (Cell Signaling technology 4695, 1:1000), anti-p-JNK (Cell Signaling technology 9255, 1:1000), anti-JNK (Cell Signaling technology 9252, 1:1000), and anti-β-actin (Abcam ab20272, 1:1000) at 4 °C overnight.

Techniques: Expressing, Activation Assay, Labeling, Staining

Journal: Nature Communications

Article Title: Sensory nerve niche regulates mesenchymal stem cell homeostasis via FGF/mTOR/autophagy axis

doi: 10.1038/s41467-023-35977-4

Figure Lengend Snippet: Sensory nerve predominates in innervating the mouse incisor, which confers the mouse incisor as an ideal model for the study of the relationship between sensory nerves and MSCs. FGF1, a ligand secreted by sensory nerves, is enriched in the proximal mesenchymal end of the incisor and surrounding Gli1 + cell. FGF1 directly regulates MSCs in the incisor by binding to FGFR1 specifically and activates the FGF/p-JNK/mTOR/autophagy axis to regulate MSCs in maintaining tissue homeostasis.

Article Snippet: Membranes were blocked with 5% non-fat dry milk for 1 h, then incubated with primary antibodies: anti-FGFR1 (Cell signaling technology 9740, 1:1000), anti-FGF1 (Abcam ab207321, 1:1000), anti-LC3 (1:1000, Abcam, ab48394), anti-p-ULK1 (Cell Signaling technology 14202, 1:1000), anti-p-mTOR (Cell Signaling technology 5536, 1:1000), anti-mTOR (Cell Signaling technology 2972, 1:1000), anti-p-p70S6K (Cell Signaling technology 9204, 1:1000), anti-p70S6K (Cell Signaling technology 2708, 1:1000), anti-p-S6 (Cell Signaling technology 4858, 1:1000), anti-S6 (Cell Signaling technology 2217, 1:1000), anti-p-Akt (Cell Signaling technology 4060, 1:1000), anti-Akt (Cell Signaling technology 9272, 1:1000), anti-p-p38 (Cell Signaling technology 4511, 1:1000), anti-p38 (Cell Signaling technology 8690, 1:1000), anti-p-ERK (Cell Signaling technology 4370, 1:1000), anti-ERK (Cell Signaling technology 4695, 1:1000), anti-p-JNK (Cell Signaling technology 9255, 1:1000), anti-JNK (Cell Signaling technology 9252, 1:1000), and anti-β-actin (Abcam ab20272, 1:1000) at 4 °C overnight.

Techniques: Binding Assay

Journal: Theranostics

Article Title: FGFR blockade boosts T cell infiltration into triple-negative breast cancer by regulating cancer-associated fibroblasts

doi: 10.7150/thno.68972

Figure Lengend Snippet: FGFR signaling pathways were enriched in immune-excluded type triple-negative breast cancer (TNBC). A) The gene signature in immune-inflamed and immune-excluded TNBC samples from TCGA dataset. B) The correlation between FGFR score and 23 types of stromal cells in TME based on TCGA BRCA dataset. “Pos.” represents immune cells positively correlated with FGFR score; “Neg.” represents immune cells negatively correlated with FGFR score; “No correlation” represents immune cells that do not correlate with FGFR score. Immune subtypes (C1-C6) were characterized by differences in the nature of the overall immune response .C) The correlation between FGFR1/2/3/4 expression and cytotoxic T lymphocytes (CTL) infiltration in indicated cancer types from GEO database based on Tumor Immune Dysfunction and Exclusion (TIDE) system. D) T cell exclusion score in BRCA of TCGA based on FGFRs expression. E) Immune phenotypes of TNBC defined by IHC staining of CD3. F) The expression of FGFR1 in immune-inflamed and immune-excluded TNBC samples based on IHC staining (inflamed, n=33; excluded, n=118, t test). G) Kaplan-Meier survival analysis of low FGFR1 (blue, n=51) versus high FGFR1 (red, n=68) expression in TNBC.

Article Snippet: Tumor specimens were stained using antibodies against α-SMA (Abcam, ab7817), CD3 (Abcam, ab5690) or FGFR1 (Cell Signaling Technology, #9740).

Techniques: Expressing, Immunohistochemistry

Journal: Theranostics

Article Title: FGFR blockade boosts T cell infiltration into triple-negative breast cancer by regulating cancer-associated fibroblasts

doi: 10.7150/thno.68972

Figure Lengend Snippet: FGFR blockade induced T cell infiltration by modulating fibroblasts. A) The effect of FGFRi Erdafitinib on CD4 + and CD8 + T cell migration was detected by transwell migration assay (n=5 biological replicates, one-way ANOVA). B) Representative staining and IHC score of α-SMA in immune-inflamed and immune-excluded TNBC samples. C) Representative IF staining of α-SMA and CD3 in immune-inflamed and immune-excluded TNBC samples. D) FGFR1 expression in tumor microenvironment of breast cancer (GSE114727). E-F) Cell population in TME of breast cancer based on FGFR1 expression. G) Representative IF staining of FGFR1 and α-SMA in TNBC samples. H) The effect of FGFRi Erdafitinib on CD4 + and CD8 + T cell migration in presence of CAFs was detected by transwell migration assay (n=3 biological replicates, one-way ANOVA).

Article Snippet: Tumor specimens were stained using antibodies against α-SMA (Abcam, ab7817), CD3 (Abcam, ab5690) or FGFR1 (Cell Signaling Technology, #9740).

Techniques: Migration, Transwell Migration Assay, Staining, Expressing

Journal: Theranostics

Article Title: FGFR blockade boosts T cell infiltration into triple-negative breast cancer by regulating cancer-associated fibroblasts

doi: 10.7150/thno.68972

Figure Lengend Snippet: FGFR blockade synergizes with immune checkpoint blockade therapy. A) Overall survival of melanoma patients who had high FGFR1 vs. low FGFR1 expressed in the tumors before anti-PD-1 treatment (GSE78220). B and C) EMT6 (B) and 4T1 (C) tumor growth in mice treated with vehicle, anti-PD-1, FGFRi (Erdafitinib) or combination of anti-PD-1 and FGFRi (n=7 mice/group, two-way ANOVA). D) Survival analysis of 4T1 tumor-bearing mice treated with indicated therapy (n=8 mice/group, log-rank test). E) The t-SNE plot of TILs and CD8 + T cell population in 4T1 tumors from mice treated with indicated therapies (n=6, one-way ANOVA). F) Percentage of IFN-γ + CD8 + T-cells in indicated therapy-treated 4T1 tumors (n=6, one-way ANOVA). G) Gene ontology (GO) analysis by RNA-seq of 4T1 tumors in indicated groups (n=3/group). Heatmap shows the DEGs and associated signatures. COM, anti-PD-1+FGFRi. H) Heatmap shows the percentage of tumor infiltrating immune cells and fibroblasts in indicated therapy-treated 4T1 tumors.

Article Snippet: Tumor specimens were stained using antibodies against α-SMA (Abcam, ab7817), CD3 (Abcam, ab5690) or FGFR1 (Cell Signaling Technology, #9740).

Techniques: RNA Sequencing Assay

Journal: Evidence-based Complementary and Alternative Medicine : eCAM

Article Title: Combination of Ligusticum Chuanxiong and Radix Paeonia Promotes Angiogenesis in Ischemic Myocardium through Notch Signalling and Mobilization of Stem Cells

doi: 10.1155/2019/7912402

Figure Lengend Snippet: Expression of angiogenesis and stem cell mobilization proteins. (a) Protein levels of HIF-1 α and FGFR-1. (b) Protein levels of SDF-1, CXCR-4, and cardiotrophin1. (c) Protein levels of Notch1 and NICD. ∗ P <0.05, ∗∗ P <0.01, and ∗∗∗ P <0.001 vs. sham group; $ P <0.05, $$ P <0.01 vs. model group; && P <0.01 vs. CX-CS group. n=3.

Article Snippet: After incubation with primary antibodies against GAPDH (5174, CST, USA), hypoxia-inducible factor 1-alpha (HIF-1 α ) (36169, CST, USA), FGF Receptor 1 (FGFR-1) (9740, CST, USA), Notch1 (3608, CST, USA), cleaved Notch1 (also named NICD) (4147s, CST), stromal cell-derived factor-1 (SDF-1) (3740, CST, USA), C-X-C chemokine receptor 4 (CXCR-4) (60042-1-1g, Proteintech, USA), and cardiotrophin1 (YT0638, ImmunoWay, USA) at 4°C, the membranes were incubated with a secondary antibody (115-035-003, Jackson, USA) at room temperature for two hours.

Techniques: Expressing

Journal: Frontiers in Pharmacology

Article Title: Deficiency of endothelial FGFR1 alleviates hyperoxia-induced bronchopulmonary dysplasia in neonatal mice

doi: 10.3389/fphar.2022.1039103

Figure Lengend Snippet: scRNA-seq analysis of lungs from normoxia- and hyperoxia-reared mice. (A) Approach to generate a single-cell atlas. (B) UMAP plot of all scRNA-seq data, showing a total of 19 distinct cell types corresponding to 5 major cell groups. Cell populations are colored as indicated by the legend. (C) Heatmap of the top 5 most differentially expressed genes across 5 major cell types. The intensity of expression is indicated as specified by the color legend. (D) Feature plots showing the expression of principal identifiers of epithelial cells, endothelial cells, stromal cells, myeloid cells, lymphocytes (B cells) and lymphocyte (T cells) populations. (E) Cellular compositions are colored as indicated by the legend in normal and hyperoxia-impaired lungs. (F) The relative proportion of endothelial cells from all cells in normal and hyperoxia-impaired lungs. (G) Hyperoxia-impacted signaling pathways in ECs, as identified by GO enrichment analysis of biological processes. (H) Violin plot showing the expression of Fgfr1 in ECs. (I) Fgfr1 + ECs in total ECs.

Article Snippet: The membranes were incubated overnight with primary anti-FGFR1 antibody (1:1000; Cell Signaling Technology, #9740).

Techniques: Expressing

Journal: Frontiers in Pharmacology

Article Title: Deficiency of endothelial FGFR1 alleviates hyperoxia-induced bronchopulmonary dysplasia in neonatal mice

doi: 10.3389/fphar.2022.1039103

Figure Lengend Snippet: RNA-seq analysis of differentially expressed genes (DEGs) and hyperoxia-impacted signaling pathways in ECs from normal and hyperoxia-impaired lungs. (A) Volcano plot showing upregulated and downregulated transcript levels of DEGs, p-value < 0.05, |log2FoldChange|≥1. (B) Heatmap of the top 50 upregulated DEGs and top 50 downregulated DEGs. (C) Hyperoxia-impacted signaling pathways in ECs as identified by GO enrichment analysis of biological processes. All terms shown are significantly enriched (p-value < 0.05). (D) Hyperoxia-impacted signaling pathways in ECs as identified by KEGG pathway enrichment analysis. All terms shown are significantly enriched (p-value < 0.05). (E) Upregulated downstream pathways of activated FGFR1 as revealed by Gene Set Enrichment Analysis, all terms shown are significantly enriched (p-value < 0.05). (F) qPCR of Fgfr1 expression in ECs of normoxic or hyperoxic lungs. n = 7 per group. Data are shown as means ± SEMs. ** p < 0.01. (G) Western blot examining the expression of FGFR1 in ECs of normoxic or hyperoxic lung, Gapdh as negative control.

Article Snippet: The membranes were incubated overnight with primary anti-FGFR1 antibody (1:1000; Cell Signaling Technology, #9740).

Techniques: RNA Sequencing Assay, Expressing, Western Blot, Negative Control

Journal: Frontiers in Pharmacology

Article Title: Deficiency of endothelial FGFR1 alleviates hyperoxia-induced bronchopulmonary dysplasia in neonatal mice

doi: 10.3389/fphar.2022.1039103

Figure Lengend Snippet: Deletion of endothelial Fgfr1 improved alveolar development and respiratory metrics and angiogenesis in mice upon hyperoxia. (A) Schematic representation of EC-specific inducible deletion of Fgfr1 in neonatal mice. (B) Representative images of H&E-stained lungs from Fgfr1 +/+ and Fgfr1 iΔEC/iΔEC mice reared in room air or hyperoxia. The top panel shows low-magnification (scale bar = 100 μm) images, and the bottom panel shows higher-magnification (scale bar = 20 μm) images. (C,D) Quantification of MLI (C) and RAC (D) based on the data in (B). Data are shown as means ± SEMs. n = 6 per group. ** p < 0.01, *** p < 0.001. (E–H) Results of respiratory metrics measurement. Data are shown as means ± SEMs. n = 9, 13 or 14 per group. * p < 0.05, ** p < 0.01, **** p < 0.0001. (I) Representative images of IF staining for VE-Cad in lung sections. Magnification: ×40. Scale bar = 20 μm. (K) Mean gray value of VE-Cad based on the data in (I). Data are shown as means ± SEMs. n = 5 per group. ** p < 0.01, *** p < 0.001. (J) Representative images of FITC-dextran leakage from mice reared in room air or hyperoxia. Magnification: ×40. Scale bar = 20 μm. (L) Mean gray value of dextran based on the data in (J). Data are shown as means ± SEMs. n = 5 per group. *** p < 0.001.

Article Snippet: The membranes were incubated overnight with primary anti-FGFR1 antibody (1:1000; Cell Signaling Technology, #9740).

Techniques: Staining

Journal: Frontiers in Pharmacology

Article Title: Deficiency of endothelial FGFR1 alleviates hyperoxia-induced bronchopulmonary dysplasia in neonatal mice

doi: 10.3389/fphar.2022.1039103

Figure Lengend Snippet: Hyperoxia induced upregulation of FGFR1 mainly in aCap cells rather than in gCap cells. (A) A total of 5 clusters of ECs were identified. Cell populations are colored as indicated by the legend. (B) Feature plots showing the expression of principal identifiers of general capillary endothelial cells (gCap), aerocyte capillary endothelial cells (aCap), arterial endothelial cells (Artery), venous endothelial cells (Vein) and lymphatic endothelial cells (Lymph). (C) Dotplot depicting the top 5 most differentially expressed genes across endothelial clusters. The intensity of expression is indicated as specified by the color legend. (D) Alteration of the relative proportion of gCap cells and aCap cells. The right panel shows the cellular compositions of ECs, colored as indicated by the legend, and the left panel shows the relative proportion of gCap cells and aCap cells from ECs. (E) Violin plots displaying the expression patterns of gCap cell and aCap cell markers. (F) Column chart showing Fgfr1 + gCap cells and Fgfr1 + aCap cells from total Fgfr1 + ECs. (G) Violin plots displaying the expression patterns of Fgfr1 in gCap cells and aCap cells. (H) qPCR of gCap cell markers, Aplnr and Kit expression and aCap cell markers, Apln and Car4 expression in ECs of lung from Fgfr1 +/+ and Fgfr1 iΔEC/iΔEC mice reared in room air or hyperoxia. n = 5 or 6 per group. Data are shown as means ± SEMs. ns no significant, * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: The membranes were incubated overnight with primary anti-FGFR1 antibody (1:1000; Cell Signaling Technology, #9740).

Techniques: Expressing

Journal: Frontiers in Pharmacology

Article Title: Deficiency of endothelial FGFR1 alleviates hyperoxia-induced bronchopulmonary dysplasia in neonatal mice

doi: 10.3389/fphar.2022.1039103

Figure Lengend Snippet: Inhibition of endothelial Fgfr1 improved alveolar development and respiratory metrics in neonatal mice in hyperoxia. (A) Representative images of H&E-stained lungs from vehicle or FGFR1 inhibitor treated mice. The top panel shows low-magnification (scale bar = 100 μm) images, and the bottom panel shows higher-magnification (scale bar = 20 μm) images. (B,C) Quantification of MLI (B) and RAC (C) based on the data in (A). Data are shown as means ± SEMs. n = 6 per group. ** p < 0.01, *** p < 0.001, **** p < 0.0001. (D–G) Results of respiratory metrics measurement. Data are shown as means ± SEMs. n = 10 per group. * p < 0.05, ** p < 0.01, **** p < 0.0001.

Article Snippet: The membranes were incubated overnight with primary anti-FGFR1 antibody (1:1000; Cell Signaling Technology, #9740).

Techniques: Inhibition, Staining