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: International Journal of Biological Sciences

Article Title: The FGFR1 Signaling Pathway Upregulates the Oncogenic Transcription Factor FOXQ1 to Promote Breast Cancer Cell Growth

doi: 10.7150/ijbs.74574

Figure Lengend Snippet: FGFR1 signaling upregulates FOXQ1 expression mainly through the ERK2-mediated downstream pathway. a. AP20187-induced FOXQ1 mRNA expression was inhibited by inhibitors of MEK or ERK1/2 in DCIS-iFGFR1 cells. DCIS-iFGFR1 cells were treated with vehicle or AP20187 in combination with 100 nM PD0325901 (a MEK inhibitor), 30 μM GDC0994 (an ERK1/2 inhibitor), 5 μM AZD5363 (an AKT inhibitor), or 2 μM GSK2110183 (an AKT inhibitor) as indicated. The relative expression level of FOXQ1 mRNA was assessed by RT-qPCR, and normalized to the expression level of β-actin mRNA. b. The effects of MEK inhibitor PD0325901 on the basal and the AP20187-induced ERK1/2 phosphorylation and FOXQ1 protein expression in DCIS-Ctrl and DCIS-iFGFR1 cells. Cells were treated with vehicle (-), AP20187, and/or PD0325901 as indicated. The indicated proteins were assayed by Western blot (WB). The average ratios of p-ERK1/2 to total ERK1/2, and FOXQ1 to PARP1 are presented in the two bar graphs. c. Inhibition of MEK or ERK1/2 blocked bFGF-induced FOXQ1 mRNA expression in MDA-MB-231 cells. Cells were treated with vehicle, bFGF, PD0325901, GDC0994, AZD5363, and/or GSK2110183 as indicated. The relative expression level of FOXQ1 mRNA was measured as described in Panel a. d. Inhibition of MEK blocked bFGF-induced FOXQ1 protein expression in MDA-MB-231 cells. Cells were treated with vehicle (-), bFGF, and/or PD0325901 as indicated. The indicated proteins were assayed by WB. The average ratios of p-ERK1/2 to total ERK1/2, and FOXQ1 to PARP1 are presented in the two bar graphs. e and f. The effects of ERK1 knockout or ERK2 knockout on the level of phosphorylated ERK2 or ERK1 and the expression levels of FOXQ1 protein and mRNA induced by FGFR1 activation. DCIS-iFGFR1 cells with ERK1 or ERK2 knockout were treated with vehicle or AP20187 (100 nM) for 3 hours. The indicated proteins were assayed by Western blot (Panel e), and the ratios of p-ERK1/2 to total ERK1/2 and FOXQ1 to GAPDH band intensities were presented. The FOXQ1 mRNA level was measured by qPCR and normalized by β-actin mRNA (Panel f). Data in all bar graphs were obtained from 3 independent experiments and presented as mean ± SD. * , ** , *** , **** , and ns, p < 0.05, 0.01, 0.001, 0.0001 and not significant, respectively were compared with the appropriate control groups by One-Way ANOVA test (Panels a, b, c, and d) or unpaired Student's t-test (Panel e and f).

Article Snippet: A primary antibody against FGFR1 ( 9740s, cell Signaling Technology ) , p - FGFR1 ( 3471, Cell Signaling Technology ) , FOXQ1 ( sc - 166264, Santa Cruz ) , c - FOS ( sc - 271243, Santa Cruz; 2250s, Cell Signaling Technology ) , ERK1 / 2 ( 9102S, Cell Signaling ) , phospho - Thr202 / Tyr204 - ERK1 / 2 ( p - ERK1 / 2 ) ( 9101S, Cell Signaling ) , β - actin ( 3700S, Cell Signaling ) , tubulin ( 2148S, Cell Signaling ) , PARP1 ( 13371 - 1 - AP, Proteintech ) , or GAPDH ( 60004 - 1 - Ig, Proteintech ) was incubated with the membrane at 4°C overnight .

Techniques: Expressing, Quantitative RT-PCR, Western Blot, Inhibition, Knock-Out, Activation Assay

Journal: International Journal of Biological Sciences

Article Title: The FGFR1 Signaling Pathway Upregulates the Oncogenic Transcription Factor FOXQ1 to Promote Breast Cancer Cell Growth

doi: 10.7150/ijbs.74574

Figure Lengend Snippet: FGFR1 signaling upregulates FOXQ1 expression mainly through the ERK2-mediated downstream pathway. a. AP20187-induced FOXQ1 mRNA expression was inhibited by inhibitors of MEK or ERK1/2 in DCIS-iFGFR1 cells. DCIS-iFGFR1 cells were treated with vehicle or AP20187 in combination with 100 nM PD0325901 (a MEK inhibitor), 30 μM GDC0994 (an ERK1/2 inhibitor), 5 μM AZD5363 (an AKT inhibitor), or 2 μM GSK2110183 (an AKT inhibitor) as indicated. The relative expression level of FOXQ1 mRNA was assessed by RT-qPCR, and normalized to the expression level of β-actin mRNA. b. The effects of MEK inhibitor PD0325901 on the basal and the AP20187-induced ERK1/2 phosphorylation and FOXQ1 protein expression in DCIS-Ctrl and DCIS-iFGFR1 cells. Cells were treated with vehicle (-), AP20187, and/or PD0325901 as indicated. The indicated proteins were assayed by Western blot (WB). The average ratios of p-ERK1/2 to total ERK1/2, and FOXQ1 to PARP1 are presented in the two bar graphs. c. Inhibition of MEK or ERK1/2 blocked bFGF-induced FOXQ1 mRNA expression in MDA-MB-231 cells. Cells were treated with vehicle, bFGF, PD0325901, GDC0994, AZD5363, and/or GSK2110183 as indicated. The relative expression level of FOXQ1 mRNA was measured as described in Panel a. d. Inhibition of MEK blocked bFGF-induced FOXQ1 protein expression in MDA-MB-231 cells. Cells were treated with vehicle (-), bFGF, and/or PD0325901 as indicated. The indicated proteins were assayed by WB. The average ratios of p-ERK1/2 to total ERK1/2, and FOXQ1 to PARP1 are presented in the two bar graphs. e and f. The effects of ERK1 knockout or ERK2 knockout on the level of phosphorylated ERK2 or ERK1 and the expression levels of FOXQ1 protein and mRNA induced by FGFR1 activation. DCIS-iFGFR1 cells with ERK1 or ERK2 knockout were treated with vehicle or AP20187 (100 nM) for 3 hours. The indicated proteins were assayed by Western blot (Panel e), and the ratios of p-ERK1/2 to total ERK1/2 and FOXQ1 to GAPDH band intensities were presented. The FOXQ1 mRNA level was measured by qPCR and normalized by β-actin mRNA (Panel f). Data in all bar graphs were obtained from 3 independent experiments and presented as mean ± SD. * , ** , *** , **** , and ns, p < 0.05, 0.01, 0.001, 0.0001 and not significant, respectively were compared with the appropriate control groups by One-Way ANOVA test (Panels a, b, c, and d) or unpaired Student's t-test (Panel e and f).

Article Snippet: A primary antibody against FGFR1 ( 9740s, cell Signaling Technology ) , p - FGFR1 ( 3471, Cell Signaling Technology ) , FOXQ1 ( sc - 166264, Santa Cruz ) , c - FOS ( sc - 271243, Santa Cruz; 2250s, Cell Signaling Technology ) , ERK1 / 2 ( 9102S, Cell Signaling ) , phospho - Thr202 / Tyr204 - ERK1 / 2 ( p - ERK1 / 2 ) ( 9101S, Cell Signaling ) , β - actin ( 3700S, Cell Signaling ) , tubulin ( 2148S, Cell Signaling ) , PARP1 ( 13371 - 1 - AP, Proteintech ) , or GAPDH ( 60004 - 1 - Ig, Proteintech ) was incubated with the membrane at 4°C overnight .

Techniques: Expressing, Quantitative RT-PCR, Western Blot, Inhibition, Knock-Out, Activation Assay

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: Incisor proximal mesenchyme from control and Gli1 CreER ;Fgfr1 fl/fl mice were collected, and tissues were incubated in RIPA buffer (Cell Signaling, 9806) with protease inhibitor (Thermo Fisher Scientific, 1861278) for 30 min at 4 °C.

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: Incisor proximal mesenchyme from control and Gli1 CreER ;Fgfr1 fl/fl mice were collected, and tissues were incubated in RIPA buffer (Cell Signaling, 9806) with protease inhibitor (Thermo Fisher Scientific, 1861278) for 30 min at 4 °C.

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: Incisor proximal mesenchyme from control and Gli1 CreER ;Fgfr1 fl/fl mice were collected, and tissues were incubated in RIPA buffer (Cell Signaling, 9806) with protease inhibitor (Thermo Fisher Scientific, 1861278) for 30 min at 4 °C.

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: Incisor proximal mesenchyme from control and Gli1 CreER ;Fgfr1 fl/fl mice were collected, and tissues were incubated in RIPA buffer (Cell Signaling, 9806) with protease inhibitor (Thermo Fisher Scientific, 1861278) for 30 min at 4 °C.

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: Incisor proximal mesenchyme from control and Gli1 CreER ;Fgfr1 fl/fl mice were collected, and tissues were incubated in RIPA buffer (Cell Signaling, 9806) with protease inhibitor (Thermo Fisher Scientific, 1861278) for 30 min at 4 °C.

Techniques: Binding Assay

Journal: Translational Lung Cancer Research

Article Title: Genome-wide epigenetic and mRNA-expression profiling followed by CRISPR/Cas9-mediated gene-disruptions corroborate the MIR141/MIR200C -ZEB1/ZEB2-FGFR1 axis in acquired EMT-associated EGFR TKI-resistance in NSCLC cells

doi: 10.21037/tlcr-22-507

Figure Lengend Snippet: Contribute of FGFR1 to EMT-E-TKI-R in HCC827 cells. (A) Distribution of FC (Log2) in mRNA expression following EMT-E-TKI-R of 56 receptor kinase genes for which RNA-sequencing data were available for HCC827 cells with EMT-E-TKI-R (HCC827EMT) relative to parental HCC827 cells (HCC827PAR). Selected receptor kinase genes are highlighted. (B) Distribution of H3K36me3 and DNA-methylation at the FGFR1 locus. For H3K36me3 is shown ChIP-sequencing enrichment values from HCC827PARand HCC827EMT. Asterisks show positions with significant difference. For DNA-methylation is shown the difference in beta-values in HCC827EMT relative to HCC827PAR cells. DMRs are indicated. (C) RT-qPCR-based mRNA expression analysis of HCC827Cas9 cells harboring control sgRNA C or FGFR1 sgRNAs F1 and F3. P0 indicates cells grown in absence of erlotinib and P2 indicates that cells were grown in presence of erlotinib for two passages. Values are normalized to expression of ACTB and subsequently normalized to the expression at P0 for sgRNA C given the value 1. In all panels SD represents one sample analyzed in technical triplicates, and * indicate changes for the given passage relative to HCC827Cas9 harboring sgRNA C with P<0.05 and FC >2. (D) Colorimetric MTS-assays showing the impact of FGFR1 depletion for cell viability using increasing concentrations of erlotinib for 72 h. Left panel shows result for FGFR1 sgRNA F1 and control sgRNA C and right panel shows result for FGFR1 sgRNA F3 and control sgRNA C. Each graph represents two independent MTS assays in where each sample was examined in technical duplicates and with SDs shown. * indicate differences for given concentrations of erlotinib with P<0.05. EMT-E-TKI-R, epithelial-mesenchymal-transition-associated EGFR tyrosine-kinase-inhibitor resistance; FC, fold change; EGFR, epidermal growth factor receptor; DMR, differential methylated region; SD, standard deviation; RT-qPCR, quantitative reverse transcription polymerase chain reaction.

Article Snippet: For western blot analysis of CRISPR/Cas9-mediated depletions the following antibodies were used: rabbit anti-ZEB1 (diluted 1/750) (Bethyl Laboratories, Montgomery, TX, USA, Cat# A301-922A, RRID:AB_1524126), rabbit anti-FGFR1 (diluted 1/500) (Cell Signaling Technology, Danvers, MA, USA, Cat# 9740, RRID:AB_11178519), rabbit anti-histone-H3 (diluted 1/10,000) (Abcam Cat# ab1791, RRID:AB_302613), mouse anti-βActin (diluted 1/5,000) (Sigma-Aldrich Cat# A5316, RRID:AB_476743), goat anti-mouse HRP conjugated (diluted 1/10,000) (Agilent, Santa Clara, CA, USA, Cat# P0447, RRID:AB_2617137), and goat anti-rabbit HRP conjugated (diluted 1/10,000) (Agilent Cat# P0448, RRID:AB_2617138).

Techniques: Expressing, RNA Sequencing Assay, DNA Methylation Assay, ChIP-sequencing, Quantitative RT-PCR, Methylation, Standard Deviation, Reverse Transcription Polymerase Chain Reaction

Journal: Journal of Cancer

Article Title: Tumor-associated inflammatory microenvironment in non-small cell lung cancer: correlation with FGFR1 and TLR4 expression via PI3K/Akt pathway

doi: 10.7150/jca.26277

Figure Lengend Snippet: Correlations between of FGFR1 and TLR4 expression and clinicopathologic features.

Article Snippet: Then, after antigen retrieval, the tissue sections were incubated with FGFR1 antibodies (monoclonal rabbit anti-human, 1:500, Cell Signaling Technology Co., Ltd, MA, USA), TLR4 antibodies (polyclonal rabbit anti-human, 1:500, Cell Signaling Technology Co., Ltd, MA, USA) at 37°C for 1 h, and then at 4°C overnight.

Techniques: Expressing, Histopathology

Journal: Journal of Cancer

Article Title: Tumor-associated inflammatory microenvironment in non-small cell lung cancer: correlation with FGFR1 and TLR4 expression via PI3K/Akt pathway

doi: 10.7150/jca.26277

Figure Lengend Snippet: Expression of FGFR1 in cancer tissues and paracancerous tissues.

Article Snippet: Then, after antigen retrieval, the tissue sections were incubated with FGFR1 antibodies (monoclonal rabbit anti-human, 1:500, Cell Signaling Technology Co., Ltd, MA, USA), TLR4 antibodies (polyclonal rabbit anti-human, 1:500, Cell Signaling Technology Co., Ltd, MA, USA) at 37°C for 1 h, and then at 4°C overnight.

Techniques: Expressing

Journal: Journal of Cancer

Article Title: Tumor-associated inflammatory microenvironment in non-small cell lung cancer: correlation with FGFR1 and TLR4 expression via PI3K/Akt pathway

doi: 10.7150/jca.26277

Figure Lengend Snippet: Immunohistochemical staining of FGFR1 and TLR4 in NSCLC tissues. No staining was detected for (A) FGFR1 and (B) TLR4 in the control group. Staining of FGFR1 (C) and TLR4 (D) in adenocarcinoma cells. Staining of FGFR1 (E) and TLR4 (F) in squamous carcinoma cells. (All photos are shown at 100 magnification).

Article Snippet: Then, after antigen retrieval, the tissue sections were incubated with FGFR1 antibodies (monoclonal rabbit anti-human, 1:500, Cell Signaling Technology Co., Ltd, MA, USA), TLR4 antibodies (polyclonal rabbit anti-human, 1:500, Cell Signaling Technology Co., Ltd, MA, USA) at 37°C for 1 h, and then at 4°C overnight.

Techniques: Immunohistochemical staining, Staining

Journal: Journal of Cancer

Article Title: Tumor-associated inflammatory microenvironment in non-small cell lung cancer: correlation with FGFR1 and TLR4 expression via PI3K/Akt pathway

doi: 10.7150/jca.26277

Figure Lengend Snippet: The relationship between FGFR1 and TLR4 expression in NSCLC.

Article Snippet: Then, after antigen retrieval, the tissue sections were incubated with FGFR1 antibodies (monoclonal rabbit anti-human, 1:500, Cell Signaling Technology Co., Ltd, MA, USA), TLR4 antibodies (polyclonal rabbit anti-human, 1:500, Cell Signaling Technology Co., Ltd, MA, USA) at 37°C for 1 h, and then at 4°C overnight.

Techniques: Expressing

Journal: Journal of Cancer

Article Title: Tumor-associated inflammatory microenvironment in non-small cell lung cancer: correlation with FGFR1 and TLR4 expression via PI3K/Akt pathway

doi: 10.7150/jca.26277

Figure Lengend Snippet: PI3K/Akt signaling is one of common pathway of the FGFR1 and TLR4 activation in NSCLC cells. A: The cells were respectively treated with culture medium (control group), TAK-242 control group, and 1ug/ml LPS for 24 hours, LPS with TAK-242 group. The expression of phosphorylated Akt was measured by Western Blot, * p < 0.05 vs the control group; ** p < 0.01 vs the control group; # p < 0.05 vs the LPS group; ## p < 0.01 vs the LPS group.B: The cells were respectively treated with culture medium (control group), BIBF1120 group, and 10ng/ml bFGF for 24 hours, bFGF with BIBF1120 group. The expression of phosphorylated Akt was measured by Western Blot, * p < 0.05 vs the control group; ** p < 0.01 vs the control group; # p < 0.05 vs the bFGF group; ## p < 0.01 vs the bFGF group.

Article Snippet: Then, after antigen retrieval, the tissue sections were incubated with FGFR1 antibodies (monoclonal rabbit anti-human, 1:500, Cell Signaling Technology Co., Ltd, MA, USA), TLR4 antibodies (polyclonal rabbit anti-human, 1:500, Cell Signaling Technology Co., Ltd, MA, USA) at 37°C for 1 h, and then at 4°C overnight.

Techniques: Activation Assay, Expressing, Western Blot

Journal: Journal of Cancer

Article Title: Tumor-associated inflammatory microenvironment in non-small cell lung cancer: correlation with FGFR1 and TLR4 expression via PI3K/Akt pathway

doi: 10.7150/jca.26277

Figure Lengend Snippet: PI3K/Akt signaling is involved in release of TNF-α and IL-6 induced by the FGFR1 and TLR4 agonists. The A549, PC-9 and SK-MES-1 (n=3) cells were respectively treated with culture medium, 1ug/ml LPS or 10ng/ml bFGF, 10µM LY294002 with or without LPS or bFGF. A:the expression of TNF-α was measured by ELISA(Mean ± SD). B: the expression of IL-6 was measured by ELISA (Mean ± SD). * p < 0.05 vs the control group, ** p < 0.01 vs the control group, # p < 0.05 vs the LPS group, ## p < 0.01 vs the LPS group, & p < 0.05 vs the bFGF group, && p < 0.01 vs the bFGF group.

Article Snippet: Then, after antigen retrieval, the tissue sections were incubated with FGFR1 antibodies (monoclonal rabbit anti-human, 1:500, Cell Signaling Technology Co., Ltd, MA, USA), TLR4 antibodies (polyclonal rabbit anti-human, 1:500, Cell Signaling Technology Co., Ltd, MA, USA) at 37°C for 1 h, and then at 4°C overnight.

Techniques: Expressing, Enzyme-linked Immunosorbent Assay

Journal: Journal of Cancer

Article Title: Tumor-associated inflammatory microenvironment in non-small cell lung cancer: correlation with FGFR1 and TLR4 expression via PI3K/Akt pathway

doi: 10.7150/jca.26277

Figure Lengend Snippet: FGFR1 regulated cell proliferation and migration by the PI3K/Akt pathway. The PC-9 cells were divided into control group, LY294002 group, bFGF group and LY294002 with bFGF group. A: the cell proliferation assay was measured by xCELLigence RTCA system. B: the the cell migration assay was measured by xCELLigence RTCA system.

Article Snippet: Then, after antigen retrieval, the tissue sections were incubated with FGFR1 antibodies (monoclonal rabbit anti-human, 1:500, Cell Signaling Technology Co., Ltd, MA, USA), TLR4 antibodies (polyclonal rabbit anti-human, 1:500, Cell Signaling Technology Co., Ltd, MA, USA) at 37°C for 1 h, and then at 4°C overnight.

Techniques: Migration, Proliferation Assay, Cell Migration Assay

Journal: International Journal of Biological Sciences

Article Title: miR-671-5p Blocks The Progression Of Human Esophageal Squamous Cell Carcinoma By Suppressing FGFR2

doi: 10.7150/ijbs.32429

Figure Lengend Snippet: FGFR2 activates the MAPK-ERK and PI3K-AKT pathways. A-E. Representative immunoblots ( A ) and plots of relative levels of FGFR2 ( B ) and p-FGFR proteins ( C ), ratios of p-ERK to ERK ( D ), ratios of p-AKT to AKT ( E ), relative levels of cyclin D1 ( F ) and cyclin B1 proteins ( G ). GAPDH serves as loading control.

Article Snippet: The following antibodies were used: anti- FGFR2, from Santa Cruz Biotechnology, anti-GAPDH from Sigma-Aldrich, and anti-AKT, anti-phosphor-AKT, anti-ERK, anti-phosphor-ERK, anti-cyclinD1, anti-cyclinB1, and anti-phosphor-FGFR from Cell Signaling Technology.

Techniques: Western Blot

Journal: International Journal of Biological Sciences

Article Title: miR-671-5p Blocks The Progression Of Human Esophageal Squamous Cell Carcinoma By Suppressing FGFR2

doi: 10.7150/ijbs.32429

Figure Lengend Snippet: miR-671-5p inhibits the phosphorylation of FGFR2 and signals of MAPK-ERK and PI3K-AKT pathways in ESCC cells . A-E. Representative immunoblots ( A ), and plots of relative levels of FGFR2 ( B ) and p-FGFR proteins ( C ), and ratios of p-AKT to AKT ( D ) and ratios of p-ERK to ERK ( E ) in cells as described in Fig. 6B .

Article Snippet: The following antibodies were used: anti- FGFR2, from Santa Cruz Biotechnology, anti-GAPDH from Sigma-Aldrich, and anti-AKT, anti-phosphor-AKT, anti-ERK, anti-phosphor-ERK, anti-cyclinD1, anti-cyclinB1, and anti-phosphor-FGFR from Cell Signaling Technology.

Techniques: Western Blot