Journal: eLife

Article Title: FGF2-FGFR1 signaling regulates release of Leukemia-Protective exosomes from bone marrow stromal cells

doi: 10.7554/elife.40033

Figure Lengend Snippet: Figure 1. Extracellular vesicles (ECVs) secreted by HS-5 cells are internalized by MOLM14 and K562 cells and protect from treatment with AC220 or imatinib, respectively. HS-5 conditioned media (CM) was collected and separated by ultracentrifugation at 100,000 g into a supernatant (S100) and pellet (P100) fraction containing ECVs. These fractions were incubated with (A) K562 cells ± 1 mM imatinib, or (B) MOLM14 cells ± 10 nM AC220, and Figure 1 continued on next page

Article Snippet: DOI: https://doi.org/10.7554/eLife.40033 14 of 23 Continued Reagent type (species) or resource Designation Source or reference Identifiers Additional information Genetic reagent (homo sapiens) FGF2-2 GenScript CRISPR/Cas nine guide RNA design Genetic reagent (homo sapiens) FGFR1-1 GenScript CRISPR/Cas nine guide RNA design Genetic reagent (homo sapiens) FGFR1-2 GenScript CRISPR/Cas nine guide RNA design Genetic reagent (mus musculus) pMIG with BCR-ABL and GFP murine retrovirus Cell line (homo sapiens) MOLM14 Dr. Yoshinobu Matsuo RRID:CVCL_7916 Cell line (homo sapiens) K562 American Type Culture Collection RRID:CVCL_0004 Cell line (homo sapiens) HS-5 Dr. Beverly Torok-Storb RRID:CVCL_3720 Cell line (homo sapiens) HS-27 Dr. Beverly Torok-Storb RRID:CVCL_0335 Antibody Mouse monoclonal anti-FGFR1 Cell Signaling 9740 Dilution 1:1000 Antibody Rabbit polyclonal anti-FGF2 Santa Cruz Sc-79 Dilution 1:500 Antibody Rabbit monoclonal anti-CD63 ABCAM ab134045 Dilution 1:1000 Antibody Rabbit polyclonal anti-CD9 Santa Cruz Sc-9148 Dilution 1:200 Antibody Mouse monoclonal anti-tsg-101 Santa Cruz Sc-7964 Dilution 1:200 Antibody Mouse monoclonal anti-actin Millipore MAB1501 Dilution 1:5000 Peptide, recombinant protein FGF2 (human) Peprotech Commercial assay or kit Thermo Scientific lentiviral transfection kit Chemical compound, drug quizartinib (AC220) LC labs Chemical compound, drug imatinib LC labs Chemical compound, drug nilotinib SelleckChem Chemical compound, drug PD173074 SelleckChem Chemical compound, drug BGJ-398 SelleckChem Chemical compound, drug doxycycline Fisher Software, algorithm CellProfiler Cell area

Techniques: Incubation

Journal: eLife

Article Title: FGF2-FGFR1 signaling regulates release of Leukemia-Protective exosomes from bone marrow stromal cells

doi: 10.7554/elife.40033

Figure Lengend Snippet: Figure 7. Fgf2 -/- mice survive significantly longer with TKI therapy in a murine BCR-ABL leukemia model. Fgf2 +/+ bone marrow was removed from donor mice and spinoculated with pMIG BCR-ABL retrovirus containing an IRES-GFP marker. The transfected bone marrow was then transplanted into lethally irradiated Fgf2 +/+ or -/- recipients. Mice were treated with 75 mg/kg/day nilotinib by oral gavage starting on day 11 of transplant. (A) Survival curves of untreated and nilotinib-treated Fgf2 +/+ and -/- mice. (B) GFP in peripheral blood was evaluated weekly and at time of euthanasia to quantify disease burden. The average GFP (percent of nucleated cells) is shown and did not differ significantly between groups indicating that all animals developed similar disease burden. Error bars indicate standard deviation. (C) Bone marrow cells from Fgf2 +/+ mice were spinoculated with pMIG BCR- ABL retrovirus containing GFP-IRES. The cells were then incubated with ECVs obtained from Fgf2 +/+ and -/- primary stroma cultured alone or with 500 nM PD173074. The next day the incubated cells were washed three times to remove cytokines and exosomes and plated in cytokine-free methylcellulose ± imatinib. After 8 days, colonies were counted and normalized to untreated condition. Graph shown on right. Error bars indicate standard error of the mean. *p<0.05 and **p<0.005. (D) Lineage-negative bone marrow cells were isolated from Fgf2 +/+ mice and cells were stained with DiO (green) tracer, washed, and immobilized on Poly-D-lysine coated chamber slides. ECVs from bone marrow stroma of Fgf2 +/+ or -/- mice were stained with DiI (red) tracer and added to the cells for a 24 hr incubation. Slides were stained with DAPI (blue) and imaged by confocal fluorescent microscopy. Movie of the z-stack images are included as Figure 7—video 1 and 2. (E) Model of bone marrow stromal FGF2 autocrine signaling and paracrine protection of leukemia cells by FGF2-containing exosomes. Figure 7 continued on next page

Article Snippet: DOI: https://doi.org/10.7554/eLife.40033 14 of 23 Continued Reagent type (species) or resource Designation Source or reference Identifiers Additional information Genetic reagent (homo sapiens) FGF2-2 GenScript CRISPR/Cas nine guide RNA design Genetic reagent (homo sapiens) FGFR1-1 GenScript CRISPR/Cas nine guide RNA design Genetic reagent (homo sapiens) FGFR1-2 GenScript CRISPR/Cas nine guide RNA design Genetic reagent (mus musculus) pMIG with BCR-ABL and GFP murine retrovirus Cell line (homo sapiens) MOLM14 Dr. Yoshinobu Matsuo RRID:CVCL_7916 Cell line (homo sapiens) K562 American Type Culture Collection RRID:CVCL_0004 Cell line (homo sapiens) HS-5 Dr. Beverly Torok-Storb RRID:CVCL_3720 Cell line (homo sapiens) HS-27 Dr. Beverly Torok-Storb RRID:CVCL_0335 Antibody Mouse monoclonal anti-FGFR1 Cell Signaling 9740 Dilution 1:1000 Antibody Rabbit polyclonal anti-FGF2 Santa Cruz Sc-79 Dilution 1:500 Antibody Rabbit monoclonal anti-CD63 ABCAM ab134045 Dilution 1:1000 Antibody Rabbit polyclonal anti-CD9 Santa Cruz Sc-9148 Dilution 1:200 Antibody Mouse monoclonal anti-tsg-101 Santa Cruz Sc-7964 Dilution 1:200 Antibody Mouse monoclonal anti-actin Millipore MAB1501 Dilution 1:5000 Peptide, recombinant protein FGF2 (human) Peprotech Commercial assay or kit Thermo Scientific lentiviral transfection kit Chemical compound, drug quizartinib (AC220) LC labs Chemical compound, drug imatinib LC labs Chemical compound, drug nilotinib SelleckChem Chemical compound, drug PD173074 SelleckChem Chemical compound, drug BGJ-398 SelleckChem Chemical compound, drug doxycycline Fisher Software, algorithm CellProfiler Cell area

Techniques: Marker, Transfection, Irradiation, Standard Deviation, Incubation, Cell Culture, Isolation, Staining, Microscopy

Journal: Respiratory Research

Article Title: PDGF-BB regulates the pulmonary vascular tone: impact of prostaglandins, calcium, MAPK- and PI3K/AKT/mTOR signalling and actin polymerisation in pulmonary veins of guinea pigs

doi: 10.1186/s12931-018-0829-5

Figure Lengend Snippet: The effect of PDGF-BB on TXB 2 and 6-keto PGF 1α. a TXB 2 -generation in dependence of the perfusion time. b Comparison of TXB 2 -generation within the groups at the same time. c 6-keto PGF 1α -generation in dependence of the perfusion time. d Comparison of 6-keto PGF 1α -generation within the groups at the same time. For all (□) control ( n = 6); (■) perfusion with PDGF-BB ( n = 6); (grey square) perfusion with imatinib / PDGF-BB ( n = 6); (□) perfusion with imatinib ( n = 6). a/c Statistics was performed by the Wilcoxon signed ranked test. b/d Statistics was performed by the Mann-Whitney U test. P < 0.05 are considered as significant: * p < 0.05 and ** p < 0.01

Article Snippet: Imatinib mesylate, amlodipine, fasudile, calphostin C, indomethacin, SC51322, PF04418948, L798106, L161982, GSK 1059615, AS 252424, 10-DEBC and SQ22536 were purchased from Tocris Bioscience (Ellisville, Missouri, USA).

Techniques: Comparison, Control, MANN-WHITNEY

Journal: Respiratory Research

Article Title: PDGF-BB regulates the pulmonary vascular tone: impact of prostaglandins, calcium, MAPK- and PI3K/AKT/mTOR signalling and actin polymerisation in pulmonary veins of guinea pigs

doi: 10.1186/s12931-018-0829-5

Figure Lengend Snippet: IPL: Effect of PDGF-BB on the pulmonary vascular tone. a Effect of PDGF-BB on P PA . b Effect of PDGF-BB on P cap. c Effect of PDGF-BB on R pre . d Effect of PDGF-BB on R post . For all: (○) control ( n = 7); (■) PDGF-BB (n = 7); (grey circle) imatinib ( n = 7); (grey square) perfused imatinib / PDGF-BB ( n = 7); (□) nebulised imatinib / PDGF-BB ( n = 6); a-d Statistics was performed by a LMM. P < 0.05 are considered as significant: * p < 0.05, ** p < 0.01 and *** p < 0.001. a grey square / grey circle Time point 0 ( § ) vs. 140 ( §§ ) min: p < 0.001. d grey circle Time point 0 ( § ) vs. 140 ( §§ ) min: p < 0.05

Article Snippet: Imatinib mesylate, amlodipine, fasudile, calphostin C, indomethacin, SC51322, PF04418948, L798106, L161982, GSK 1059615, AS 252424, 10-DEBC and SQ22536 were purchased from Tocris Bioscience (Ellisville, Missouri, USA).

Techniques: Control

Journal: Frontiers in Genetics

Article Title: Inhibition of ABL1 by tyrosine kinase inhibitors leads to a downregulation of MLH1 by Hsp70-mediated lysosomal protein degradation

doi: 10.3389/fgene.2022.940073

Figure Lengend Snippet: MLH1 protein levels are reduced in SW480 colorectal cells after treatment with Dasatinib and imatinib/nilotnib tyrosine kinase inhibitors. (A) Western blot analysis and quantification of MLH1 protein expression after treatment with Dasatinib in SW480 colorectal cancer cells n = 3 statistical significance was determined by unpaired t-test* p < 0.05 (B) Western blot analysis and quantification of MLH1 protein expression after treatment with imatinib (top) and nilotinib (bottom) in SW480 colorectal cancer cells n = 3 statistical significance was determined by 1-way ANOVA with Bonferroni post-test n = 3 * p < 0.05, ** p < 0.01, *** p < 0.001 (C) Target comparison of Dasatinib versus imatinib/nilotinib based on .

Article Snippet: Imatinib mesylate (imatinib) was obtained from Tocris Biosciences; 10 mM stock was prepared in dimethylsulfoxide (DMSO) (Fisher Scientific) and stored at −20°C.

Techniques: Western Blot, Expressing, Comparison

Journal: Frontiers in Genetics

Article Title: Inhibition of ABL1 by tyrosine kinase inhibitors leads to a downregulation of MLH1 by Hsp70-mediated lysosomal protein degradation

doi: 10.3389/fgene.2022.940073

Figure Lengend Snippet: MLH1 protein levels are decreased and MMR function inhibited in HEK293 cells after treatment with tyrosine kinase inhibitors. (A) Western blot analysis and quantification of MLH1 protein expression after treatment with imatinib (top) and nilotinib (bottom) in HEK293 cells statistical significance determined by 1-way ANOVA with Bonferroni post-test n = 3 * p < 0.05 (B) Treatment with imatinib or nilotinib alone does not significantly affect cell viability in HEK293 cells measured via cell count 24 h after treatment (C) Cell survival measured via viable cell count after 48-h co-treatment with imatinib/nilotinib and 6TG, statistical significance was determined by unpaired t-test n = 3 * p < 0.05.

Article Snippet: Imatinib mesylate (imatinib) was obtained from Tocris Biosciences; 10 mM stock was prepared in dimethylsulfoxide (DMSO) (Fisher Scientific) and stored at −20°C.

Techniques: Western Blot, Expressing, Cell Counting

Journal: Frontiers in Genetics

Article Title: Inhibition of ABL1 by tyrosine kinase inhibitors leads to a downregulation of MLH1 by Hsp70-mediated lysosomal protein degradation

doi: 10.3389/fgene.2022.940073

Figure Lengend Snippet: ABL1 phosphorylates MLH1. (A) MLH1 mRNA fold change with ABL1 knockdown (left) or inhibition by imatinib (right) determined by RT-qPCR n = 3 (B) MLH1 immunoprecipitation in HEK293 cells with ABL1 and MLH1 overexpressed followed by immunoblot with anti-phosphotyrosine antibody. Treatment with 5 μM nilotinib reduces the phospho-tyrosine signal associated with MLH1. (C) Kinase assay using recombinant ABL1 (25 ng) and MLH1 (100 ng) protein incubated for 15 min, followed by SDS-PAGE and immunoblot analysis indicates tyrosine phosphorylation of MLH1 only in the presence of ABL1 kinase.

Article Snippet: Imatinib mesylate (imatinib) was obtained from Tocris Biosciences; 10 mM stock was prepared in dimethylsulfoxide (DMSO) (Fisher Scientific) and stored at −20°C.

Techniques: Knockdown, Inhibition, Quantitative RT-PCR, Immunoprecipitation, Western Blot, Kinase Assay, Recombinant, Incubation, SDS Page, Phospho-proteomics

Journal: Cancer Discovery

Article Title: Combined Inhibition of MAP Kinase and KIT Signaling Synergistically Destabilizes ETV1 and Suppresses GIST Tumor Growth

doi: 10.1158/2159-8290.cd-14-0985

Figure Lengend Snippet: Figure 3. ETV1 positively regulates Kit expression in murine GISTs. A, heatmap of signifi cantly differentially expressed genes between corn oil control– and tamoxifen-treated murine GIST tumors identifi ed by RNA-seq. Clustering was based on the most differentially expressed 228 genes with FDR <0.05 and fold change >2.0. Samples are color coded based on treatment status: pink, corn oil–treated; orange, tamoxifen-treated. Scale bar, mean normalized fold change by log 2 . B, RNA-seq gene expression quantifi cation (FPKM, fragments per kilobase mapped) of Kit and a representative group of ETV1 transcriptional targets in tamoxifen-treated versus corn oil–treated murine GISTs. C, representative IF images of ETV1 (green) and KIT (red) protein in cecal tumors from Etv1 fl ox/fl ox ;Kit V558Δ/+ ; Rosa26 CreERT2/CreERT2 mice treated with tamoxifen or corn oil, demonstrating ETV1 ablation and decreased KIT protein level. Nuclei (DAPI, blue). Scale bar, 50 μm. D, representative KIT IHC images of the cecal tumors and ICC hyperplasia in the large intestines of mice treated as in C. Scale bars, 50 μm. E, GSEA plots of the ranked list of the differentially expressed genes between tamoxifen (Tam)-treated versus corn oil–treated murine GIST tumor samples, using two gene sets, Imatinib UP (imatinib upregulated) and Imatinib DN (imatinib downregulated). F, GSEA plots of the ranked list of the differen- tially expressed genes between tamoxifen-treated versus corn oil–treated murine GIST tumor samples, using the ISHIDA_E2F_TARGETS gene set. GEMM, genetically engineered mouse model; NES, normalized enrichment score.

Article Snippet: Vehicle ImatinibMEK162 Imatinib+ MEK162 K i6 7 T ric hr om e % o f K i6 7 st ai ni ng 0 Ve hic le ME K1 62 Ima tini b Ima tini b+ ME K1 62 Ve hic le ME K1 62 Ima tini b Ima tini b+ ME K1 62 10 20 30 40 P = 0.004 P < 0.0001 P < 0.0001 P = 0.005 A T um or w ei gh t ( m g) 0 200 400 600 800 1,000 ns P = 0.03 P = 0.01 P = 0.04 B D MEK162 Imatinib Imatinib + MEK162Vehicle pKIT pERK ETV1 ERK KIT Actin DUSP6 GIST GEMM (Kit Δ558/+) C MARCH 2015 CANCER DISCOVERY | 313 Abcam; ab8895), rabbit anti-H3K4me3(for ChIP; Abcam; ab8580), rabbit anti-phosphorylated KIT (Cell Signaling Technology; #3073; 1:1,000 for Western blotting), rabbit anti-phosphorylated ERK1/2 (Cell Signaling Technology; #4370; 1:5,000 for Western blotting, 1:400 for IHC), rabbit anti-ERK1/2 (Cell Signaling Technology; #4695; 1:5,000 for Western blotting), rabbit anti-phosphorylated AKT (Cell Signaling Technology; #4060; 1:1,000 for Western blotting), rabbit AKT (Cell Signaling Technology; #4685; 1:1,000 for Western blotting), rabbit anti-phosphorylated S6 (Cell Signaling Technology; #2317; 1:1,000 for Western blotting, 1:400 for IHC), rabbit anti-S6 (Cell Signaling Technology; #4856; 1:1,000 for Western blotting), rabbit anti-cleaved caspase-3 (Cell Signaling Technology; #9661; 1:1,000 for Western blotting, 1:400 for IHC), horseradish peroxidase (HRP)–conjugated anti-GAPDH (Abcam; ab9385; 1:5,000 for Western blotting), HRP-conjugated anti-actin (Abcam; ab49900; 1:5,000 for Western blotting).

Techniques: Expressing, Control, RNA Sequencing, Gene Expression

Journal: Cancer Discovery

Article Title: Combined Inhibition of MAP Kinase and KIT Signaling Synergistically Destabilizes ETV1 and Suppresses GIST Tumor Growth

doi: 10.1158/2159-8290.cd-14-0985

Figure Lengend Snippet: Figure 5. Combined inhibition of MAP kinase and KIT signaling destabilizes ETV1 protein and results in enhanced growth suppression of human GIST cells. A, immunoblot of ETV1, pKIT, and pERK levels in GIST882 and GIST-T1 cells treated with imatinib (500 nmol/L) or MEK162 (1 μmol/L) for the indi- cated time points. B, ETV1 localization at the target gene loci (i.e., KIT and DUSP6 ) by ChIP–qPCR in GIST cells treated with imatinib (1 μmol/L) or MEK162 (500 nmol/L) for 8 hours in GIST882 cells, or imatinib (80 nmol/L) or MEK162 (500 nmol/L) for 2 hours in GIST-T1 cells. C, immunoblot of ETV1 and KIT, MAP kinase, and AKT signaling pathways in GIST882 and GIST-T1 cells treated with various doses of imatinib and MEK162 as indicated for 8 hours. D, cell viability by Alamar Blue of GIST882 and GIST-T1 cells treated with various doses of imatinib and MEK162 as indicated for 7 days. n = 3, mean ± SEM. E, cell viability by Alamar Blue of GIST-T1 cell expressing different MEK constructs treated with various doses of imatinib and MEK162 as indicated for 7 days. n = 3, mean ± SEM. F, immunoblot of ETV1, KIT, and MAP kinase signaling in GIST-T1 parental cells, GIST-T1 cells expressing MEK1 WT , MEK1 L115P , MEK2 WT , and MEK2 L119P . Cells were treated for 1 hour as indicated. V, DMSO; I, imatinib (500 nmol/L); M, MEK162 (1,000 nmol/L).

Article Snippet: Vehicle ImatinibMEK162 Imatinib+ MEK162 K i6 7 T ric hr om e % o f K i6 7 st ai ni ng 0 Ve hic le ME K1 62 Ima tini b Ima tini b+ ME K1 62 Ve hic le ME K1 62 Ima tini b Ima tini b+ ME K1 62 10 20 30 40 P = 0.004 P < 0.0001 P < 0.0001 P = 0.005 A T um or w ei gh t ( m g) 0 200 400 600 800 1,000 ns P = 0.03 P = 0.01 P = 0.04 B D MEK162 Imatinib Imatinib + MEK162Vehicle pKIT pERK ETV1 ERK KIT Actin DUSP6 GIST GEMM (Kit Δ558/+) C MARCH 2015 CANCER DISCOVERY | 313 Abcam; ab8895), rabbit anti-H3K4me3(for ChIP; Abcam; ab8580), rabbit anti-phosphorylated KIT (Cell Signaling Technology; #3073; 1:1,000 for Western blotting), rabbit anti-phosphorylated ERK1/2 (Cell Signaling Technology; #4370; 1:5,000 for Western blotting, 1:400 for IHC), rabbit anti-ERK1/2 (Cell Signaling Technology; #4695; 1:5,000 for Western blotting), rabbit anti-phosphorylated AKT (Cell Signaling Technology; #4060; 1:1,000 for Western blotting), rabbit AKT (Cell Signaling Technology; #4685; 1:1,000 for Western blotting), rabbit anti-phosphorylated S6 (Cell Signaling Technology; #2317; 1:1,000 for Western blotting, 1:400 for IHC), rabbit anti-S6 (Cell Signaling Technology; #4856; 1:1,000 for Western blotting), rabbit anti-cleaved caspase-3 (Cell Signaling Technology; #9661; 1:1,000 for Western blotting, 1:400 for IHC), horseradish peroxidase (HRP)–conjugated anti-GAPDH (Abcam; ab9385; 1:5,000 for Western blotting), HRP-conjugated anti-actin (Abcam; ab49900; 1:5,000 for Western blotting).

Techniques: Inhibition, Western Blot, ChIP-qPCR, Protein-Protein interactions, Expressing, Construct

Journal: Cancer Discovery

Article Title: Combined Inhibition of MAP Kinase and KIT Signaling Synergistically Destabilizes ETV1 and Suppresses GIST Tumor Growth

doi: 10.1158/2159-8290.cd-14-0985

Figure Lengend Snippet: Figure 6. Combined inhibition of the MAP kinase and KIT signaling synergistically suppresses tumor growth in in vivo GIST xenograft mouse models. A, treatment response of GIST882 xenografts in SCID mice. The treatment cohorts are as follows: (i) Vehicle (blue): water; (ii) imatinib (green): 100 mg/kg twice a day; (iii) MEK162 (red): 30 mg/kg twice a day; (iv) imatinib + MEK162 (dose 1; magenta): imatinib (100 mg/kg twice a day) + MEK162 (10 mg/kg twice a day); (v) imatinib + MEK162 (dose 2; yellow): imatinib (50 mg/kg twice a day) + MEK162 (30 mg/kg twice a day); (vi) imatinib + MEK162 (dose 3; black): imatinib (100 mg/kg twice a day) + MEK162 (30 mg/kg twice a day; dose 3; black). n = 6–8, mean ± SEM. Two-tailed unpaired t test: *, P < 0.0001; **, P < 0.0001; ***, P < 0.0001. B, representative H&E images of GIST882 xenografts after 14 days of drug treatment by oral gavage as indicated. Vehicle: water; imatinib: 100 mg/kg twice a day; MEK162: 30 mg/kg twice a day; imatinib (100 mg/kg twice a day) + MEK162 (30 mg/kg twice a day). Scale bar, 50 μm. C, immunoblots of three representative GIST882 xenograft tumors explanted after 2 days of drug treatment by oral gavage as indicated. Vehicle: water; imatinib: 100 mg/kg twice a day; MEK162: 30 mg/kg twice a day; imatinib (100 mg/kg twice a day) + MEK162 (30 mg/kg twice a day). D, treatment response of GIST-T1 xenografts in SCID mice as indicated by oral gavage. The treatment cohorts are as follows: (i) Vehicle: water; (ii) imatinib: 80 mg/kg twice a day; (iii) MEK162: 30 mg/kg twice a day; (iv) imatinib (80 mg/kg twice a day) + MEK162 (30 mg/kg twice a day). n = 10, mean ± SEM. Two-tailed unpaired t test: *, P < 0.0001; **, P < 0.0001; ***, P < 0.0001. E, representative H&E images of GIST-T1 xenografts after 21 days of drug treatment by oral gavage as indicated. Vehicle (blue): water; imatinib (green): 80 mg/kg twice a day; MEK162 (red): 30 mg/kg twice a day; imatinib + MEK162 (magenta): imatinib (80 mg/kg twice a day) + MEK162 (30 mg/kg twice a day). Scale bar, 50 μm. F, immunoblots of three representative GIST-T1 xenograft tumors explanted after 2 days of drug treatment by oral gavage as indicated. Vehicle: water; imatinib: 80 mg/kg twice a day; MEK162: 30 mg/kg twice a day; imatinib (80 mg/kg twice a day) + MEK162 (30 mg/kg twice a day).

Article Snippet: Vehicle ImatinibMEK162 Imatinib+ MEK162 K i6 7 T ric hr om e % o f K i6 7 st ai ni ng 0 Ve hic le ME K1 62 Ima tini b Ima tini b+ ME K1 62 Ve hic le ME K1 62 Ima tini b Ima tini b+ ME K1 62 10 20 30 40 P = 0.004 P < 0.0001 P < 0.0001 P = 0.005 A T um or w ei gh t ( m g) 0 200 400 600 800 1,000 ns P = 0.03 P = 0.01 P = 0.04 B D MEK162 Imatinib Imatinib + MEK162Vehicle pKIT pERK ETV1 ERK KIT Actin DUSP6 GIST GEMM (Kit Δ558/+) C MARCH 2015 CANCER DISCOVERY | 313 Abcam; ab8895), rabbit anti-H3K4me3(for ChIP; Abcam; ab8580), rabbit anti-phosphorylated KIT (Cell Signaling Technology; #3073; 1:1,000 for Western blotting), rabbit anti-phosphorylated ERK1/2 (Cell Signaling Technology; #4370; 1:5,000 for Western blotting, 1:400 for IHC), rabbit anti-ERK1/2 (Cell Signaling Technology; #4695; 1:5,000 for Western blotting), rabbit anti-phosphorylated AKT (Cell Signaling Technology; #4060; 1:1,000 for Western blotting), rabbit AKT (Cell Signaling Technology; #4685; 1:1,000 for Western blotting), rabbit anti-phosphorylated S6 (Cell Signaling Technology; #2317; 1:1,000 for Western blotting, 1:400 for IHC), rabbit anti-S6 (Cell Signaling Technology; #4856; 1:1,000 for Western blotting), rabbit anti-cleaved caspase-3 (Cell Signaling Technology; #9661; 1:1,000 for Western blotting, 1:400 for IHC), horseradish peroxidase (HRP)–conjugated anti-GAPDH (Abcam; ab9385; 1:5,000 for Western blotting), HRP-conjugated anti-actin (Abcam; ab49900; 1:5,000 for Western blotting).

Techniques: Inhibition, In Vivo, Two Tailed Test, Western Blot

Journal: BioMed Research International

Article Title: Pharmacokinetics Interaction between Imatinib and Genistein in Rats

doi: 10.1155/2015/368976

Figure Lengend Snippet: Extracted ion chromatograms for (a) blank rat plasma, (b) blank rat plasma spiked with imatinib (10 μg/mL), N-desmethyl imatinib (0.500 μg/mL), and IS (carbamazepine), and (c) rat plasma sample after oral administration of single dosage 30 mg/kg imatinib. (1) Imatinib, (2) N-desmethyl imatinib, and (3) IS (carbamazepine).

Article Snippet: N-Desmethyl imatinib (purity > 98%) was purchased from Toronto Research Chemicals Inc. (North York, Canada).

Techniques: Clinical Proteomics

Journal: BioMed Research International

Article Title: Pharmacokinetics Interaction between Imatinib and Genistein in Rats

doi: 10.1155/2015/368976

Figure Lengend Snippet: The main pharmacokinetic parameters of imatinib in five groups ( n = 5).

Article Snippet: N-Desmethyl imatinib (purity > 98%) was purchased from Toronto Research Chemicals Inc. (North York, Canada).

Techniques:

Journal: BioMed Research International

Article Title: Pharmacokinetics Interaction between Imatinib and Genistein in Rats

doi: 10.1155/2015/368976

Figure Lengend Snippet: Mean concentration-time curve of imatinib in five groups ( n = 5).

Article Snippet: N-Desmethyl imatinib (purity > 98%) was purchased from Toronto Research Chemicals Inc. (North York, Canada).

Techniques: Concentration Assay

Journal: BioMed Research International

Article Title: Pharmacokinetics Interaction between Imatinib and Genistein in Rats

doi: 10.1155/2015/368976

Figure Lengend Snippet: Mean concentration-time curve of N-desmethyl imatinib in five groups ( n = 5).

Article Snippet: N-Desmethyl imatinib (purity > 98%) was purchased from Toronto Research Chemicals Inc. (North York, Canada).

Techniques: Concentration Assay

Journal: BioMed Research International

Article Title: Pharmacokinetics Interaction between Imatinib and Genistein in Rats

doi: 10.1155/2015/368976

Figure Lengend Snippet: The main pharmacokinetic parameters of N-desmethyl imatinib in five groups ( n = 5).

Article Snippet: N-Desmethyl imatinib (purity > 98%) was purchased from Toronto Research Chemicals Inc. (North York, Canada).

Techniques:

Journal: bioRxiv

Article Title: Loss of endothelial ALK1 signaling induces the emergence of a KIT+ angiogenic endothelial cluster driving brain arteriovenous malformations

doi: 10.1101/2025.06.05.657957

Figure Lengend Snippet: A , Experimental timeline of tamoxifen and KIT inhibitor administration in Alk1iECKO and control mice. B, D, F , CD31 immunostaining of the PNVP of P8 Alk1iECKO mice injected with imatinib ( B ), masitinib ( D ), and KIT blocking antibody ( F ), along with their corresponding vehicle controls. C, E, G , Quantification of vessel diameter in the PNVP of P8 Alk1iECKO mice injected with the indicated inhibitor. Each dot represents one mouse. Error bars represent means ± s.e.m, *P<0.05, **P<0.01, ***P<0.001, Mann-Whitney test, Welch’s t test or Unpaired t test (B, D, F) were performed.

Article Snippet: Imatinib Mesylate (100 mg/kg, Selleckchem, S1026), Masitinib Mesylate (100mg/Kg, Cerdalane, E0814), KIT blocking antibody (500 µg, InVivo MAb anti-mouse c-Kit, BioxCell, BE0293-5MG-A) were administered intraperitoneally at P6, 3 hours following tamoxifen injection, and again at P7.

Techniques: Control, Immunostaining, Injection, Blocking Assay, MANN-WHITNEY

Journal: bioRxiv

Article Title: Loss of endothelial ALK1 signaling induces the emergence of a KIT+ angiogenic endothelial cluster driving brain arteriovenous malformations

doi: 10.1101/2025.06.05.657957

Figure Lengend Snippet: A , C , CD31 immunostaining of the PNVP of P8 Alk1l/l mice injected with imatinib (A), and masitinib (C), along with their corresponding vehicle controls. B, D , Quantification of vessel diameter in the PNVP of P8 Alk1l/l mice injected with the indicated inhibitor. Each dot represents one mouse. Error bars represent means ± s.e.m, Mann-Whitney test or Unpaired t test (B, D) were performed. D , Schematic model illustrating how 48 hours of Alk1 deletion induces the emergence of angiogenic 1 ECs in both PNVP and INVP. In the PNVP, angiogenic 1 ECs further differentiate into angiogenic 2 ECs, which drive AVM formation. Both angiogenic populations express KIT, with angiogenic 2 showing stronger expression. Pharmacological inhibition of KIT effectively prevents AVM development in this model. Each dot represents one mouse.

Article Snippet: Imatinib Mesylate (100 mg/kg, Selleckchem, S1026), Masitinib Mesylate (100mg/Kg, Cerdalane, E0814), KIT blocking antibody (500 µg, InVivo MAb anti-mouse c-Kit, BioxCell, BE0293-5MG-A) were administered intraperitoneally at P6, 3 hours following tamoxifen injection, and again at P7.

Techniques: Immunostaining, Injection, MANN-WHITNEY, Expressing, Inhibition