Journal: Gastric Cancer

Article Title: Targeting yes-associated protein to overcome imatinib resistance in gastrointestinal stromal tumor drug-tolerant persister cells

doi: 10.1007/s10120-025-01657-z

Figure Lengend Snippet: Characterization of imatinib-derived gastrointestinal stromal tumor (GIST) drug-tolerant persister cells (DTPs). A Schematic explanation of DTP generation. B Cell proliferation measured by WST-8 assays at 72 h after imatinib treatment in GIST-T1 parental cells and DTPs. Each value is presented as mean ± SD (n = 6). Two-sided t-test (*** p < 0.001). C Western blot analysis of cKIT, pKIT, YAP and pYAP in nuclear and whole cell lysates of parental cells, DTPs, and regrown cells. D Cell cycle analysis of parental cells and DTPs. E Immunofluorescence staining of YAP/TAZ in parental cells, DTPs, and regrown cells. Scale bar:20 μm

Article Snippet: Cells were treated with imatinib mesylate (Tocris, Bristol, UK), verteporfin (a YAP inhibitor (Selleck chemicals, Houston, TX), or XAV-939 (a tankyrase inhibitor that indirectly suppresses YAP activity; Selleck chemicals, Houston, TX) [ ].

Techniques: Derivative Assay, Western Blot, Cell Cycle Assay, Immunofluorescence, Staining

Journal: Gastric Cancer

Article Title: Targeting yes-associated protein to overcome imatinib resistance in gastrointestinal stromal tumor drug-tolerant persister cells

doi: 10.1007/s10120-025-01657-z

Figure Lengend Snippet: Evaluation of YAP in patient samples A immunohistochemistry (IHC) staining for YAP in GIST patient samples with or without preoperative imatinib treatment. Scale bar:50 μm. B Proportion of YAP-positive tumor cell nuclei in 26 untreated or imatinib-resistant cases and 17 imatinib-responsive cases. Each value is presented as mean ± SEM. Mann–Whitney U test (**p < 0.01)

Article Snippet: Cells were treated with imatinib mesylate (Tocris, Bristol, UK), verteporfin (a YAP inhibitor (Selleck chemicals, Houston, TX), or XAV-939 (a tankyrase inhibitor that indirectly suppresses YAP activity; Selleck chemicals, Houston, TX) [ ].

Techniques: Immunohistochemistry, MANN-WHITNEY

Journal: Gastric Cancer

Article Title: Targeting yes-associated protein to overcome imatinib resistance in gastrointestinal stromal tumor drug-tolerant persister cells

doi: 10.1007/s10120-025-01657-z

Figure Lengend Snippet: Effects of YAP inhibitors on DTPs. A Western blot analysis of YAP and pYAP in nuclear and whole-cell lysates of parental cells, DTPs, and DTPs treated with verteporfin or XAV-939. B Immunofluorescence staining of YAP/TAZ in DTPs treated with verteporfin or XAV-939. Scale bar:20 μm. C Cell proliferation measured by WST-8 assays at 72 h after verteporfin or XAV-939 treatment in parental cells and DTPs. Each value is presented as mean ± SD (n = 6). Two-sided t-test (* p < 0.05, ** p < 0.01, *** p < 0.001). ( D ) Caspase 3/7 assay of the DTPs at 8 h after treatment with DMSO, imatinib, verteporfin or XAV-939, respectively. Each value is presented as mean ± SD (n = 4). Two-sided t-test (* p < 0.05)

Article Snippet: Cells were treated with imatinib mesylate (Tocris, Bristol, UK), verteporfin (a YAP inhibitor (Selleck chemicals, Houston, TX), or XAV-939 (a tankyrase inhibitor that indirectly suppresses YAP activity; Selleck chemicals, Houston, TX) [ ].

Techniques: Western Blot, Immunofluorescence, Staining

Journal: Gastric Cancer

Article Title: Targeting yes-associated protein to overcome imatinib resistance in gastrointestinal stromal tumor drug-tolerant persister cells

doi: 10.1007/s10120-025-01657-z

Figure Lengend Snippet: Effects of combination therapies of imatinib and YAP inhibitors. A Proliferation of cells treated with the following therapies, followed by drug washout: (1) vehicle, (2) verteporfin for 3 d, (3) XAV-939 for 3 d, (4) imatinib for 12 d, (5) imatinib for 9 d followed by a combination of imatinib and verteporfin for 3 d, and (6) imatinib for 9 d followed by a combination of imatinib and XAV-939 for 3 d. Each value is presented as mean ± SD (n = 6). Tukey–Kramer honestly significant difference test (** p < 0.01, *** p < 0.001). B Cell cycle assay of DTPs treated with the indicated therapies and incubated for 8 d after drug washout: (1) imatinib for 3 d, (2) imatinib and verteporfin for 3 d, and (3) imatinib and XAV-939 for 3 d. Each value is presented as mean ± SD (n = 3). Two-sided t-test (*** p < 0.001). C Annexin V-FITC/DAPI apoptosis assay in parental cells and DTPs treated with the indicated therapies for 8 h. Parental cells: (1) vehicle, (2) imatinib, (3) verteporfin, (4) XAV-939. DTPs: (5) vehicle, (6) imatinib, (7) imatinib and verteporfin, and (8) imatinib and XAV-939

Article Snippet: Cells were treated with imatinib mesylate (Tocris, Bristol, UK), verteporfin (a YAP inhibitor (Selleck chemicals, Houston, TX), or XAV-939 (a tankyrase inhibitor that indirectly suppresses YAP activity; Selleck chemicals, Houston, TX) [ ].

Techniques: Cell Cycle Assay, Incubation, Apoptosis Assay

Journal: Gastric Cancer

Article Title: Targeting yes-associated protein to overcome imatinib resistance in gastrointestinal stromal tumor drug-tolerant persister cells

doi: 10.1007/s10120-025-01657-z

Figure Lengend Snippet: Antitumor effects of verteporfin in a GIST xenograft mouse model. A GIST-T1 cells were transplanted subcutaneously and treated with the following therapies: (1) vehicle (n = 4), (2) verteporfin for 3 d (n = 4), (3) imatinib for 12 d (n = 4), and (4) imatinib for 9 d followed by a combination of imatinib and verteporfin for 3 d (n = 4). Treatment was initiated when tumor volume reached approximately 500 mm 3 (day 0). B Body weight and ( C ) tumor volume were measured every day until day 35. Each value is presented as mean ± SD (n = 4). Two-sided t-test (* p < 0.05, ** p < 0.01, *** p < 0.001). D Representative YAP-stained tumor sections from each treatment group. Scale bar: 25 μm. E Proportion of YAP positive nuclei. Each value is presented as mean ± SEM (n = 3), Tukey–Kramer honestly significant difference test (* p < 0.05, ** p < 0.01). F Apoptosis analysis by TUNEL staining from each treatment group. Scale bar: 100 µm

Article Snippet: Cells were treated with imatinib mesylate (Tocris, Bristol, UK), verteporfin (a YAP inhibitor (Selleck chemicals, Houston, TX), or XAV-939 (a tankyrase inhibitor that indirectly suppresses YAP activity; Selleck chemicals, Houston, TX) [ ].

Techniques: Staining, TUNEL Assay

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

Journal: Frontiers in Pharmacology

Article Title: c-Abl Inhibition Exerts Symptomatic Antiparkinsonian Effects Through a Striatal Postsynaptic Mechanism

doi: 10.3389/fphar.2018.01311

Figure Lengend Snippet: Effects of imatinib and levodopa on striatal motor behaviors and c-Abl/Cdk5/DARPP-32 signaling cascades. (A) Striatal penetration of intraperitoneally injected imatinib in mice. HPLC analysis were done to quantify concentrations of imatinib in the striatum ( n = 4), cortex ( n = 5), hippocampus ( n = 4), thalamus ( n = 4), and blood plasma ( n = 5) of naïve mice that received single i.p. injections of imatinib mesylate (25 mg/kg) 30 min before sacrifice. Values are expressed as means ± SEM. (B) Symptomatic antiparkinsonian effects of imatinib and levodopa in MPTP-treated mice. Behavioral tests were carried out in vehicle or MPTP-treated mice 30 min after a single i.p. injection of imatinib and/or levodopa. ( left-upper panel ) The beam-walking test for examining the effects of administration of imatinib mesylate (25 mg/kg) or levodopa (15 mg/kg). Values are means ± SEM ( n = 5–21). ∗ P < 0.05 versus vehicle-treated mice; one-way ANOVA [ F (5,77) = 11.265] followed by the Scheffe post hoc test. ( right-upper panel ) The rota-rod test for examining the effects of imatinib mesylate (25 mg/kg) or levodopa (15 mg/kg) administration. Values are means ± SEM ( n = 8–21). ∗ P < 0.05 versus vehicle-treated mice; one-way ANOVA [ F (5,80) = 7.710] followed by the Scheffe post hoc test. ( left-lower panel ) The beam-walking test for examining the effects of imatinib mesylate (10 mg/kg) and/or levodopa (2.5 or 5 mg/kg) administration. Values are means ± SEM ( n = 10–11). # P < 0.05 versus MPTP-treated mice; one-way ANOVA [ F (5,55) = 4.177] followed by the Scheffe post hoc test. ( right-lower panel ) The rota-rod test for examining the effects of imatinib mesylate (10 mg/kg) and/or levodopa (2.5 or 5 mg/kg) administration. Values are means ± SEM ( n = 10–11). ### P < 0.001 versus MPTP-treated mice; one-way ANOVA [ F (5,55) = 8.283] followed by the Scheffe post hoc test. (C) Western-blot analysis of striatal levels of Cdk5-pTyr15 and Cdk5 in vehicle or MPTP-treated mice 30 min after single i.p. injections of imatinib. Values are means ± SEM ( n = 4–5). ∗ P < 0.05 versus vehicle-treated mice, # P < 0.05 versus MPTP-treated mice; one-way ANOVA [ F Cdk5-pTyr15(4,19) = 50.391, F Cdk5(4,19) = 1.413] followed by the Scheffe post hoc test. IMB (10), imatinib mesylate (10 mg/kg); IMB (25), imatinib mesylate (25 mg/kg). (D) Western-blot analysis of striatal levels of DARPP-32-pThr75, DARPP-32-pThr34, and DARPP-32 in vehicle or MPTP-treated mice 30 min after a single i.p. injection of imatinib. Values are means ± SEM ( n = 4–5). ∗ P < 0.05 versus vehicle-treated mice, # P < 0.05 versus MPTP-treated mice; one-way ANOVA [ F DARPP-32-pThr75(4,19) = 35.089, F DARPP-32-pThr34(4,19) = 0.711, F DARPP-32(4,19) = 0.293] followed by the Scheffe post hoc test. IMB (10), imatinib mesylate (10 mg/kg); IMB (25), imatinib mesylate (25 mg/kg). (E) Western-blot analysis of striatal levels of Cdk5-pTyr15 and Cdk5 in MPTP-treated mice 30 min after a single i.p. injection of levodopa and/or imatinib. Values are expressed as means ± SEM ( n = 5–10). # P < 0.05, ## P < 0.01 versus MPTP-treated mice. One-way ANOVA [ F Cdk5-pTyr15(3,31) = 6.039, F Cdk5(3,17) = 0.258] followed by the Scheffe post hoc test. Levodopa (5), levodopa (5 mg/kg); IMB (10), imatinib mesylate (10 mg/kg). (F) Western-blot analysis of striatal levels of DARPP-32-pThr75, DARPP-32-pThr34, and DARPP-32 in MPTP-treated mice 30 min after a single i.p. injection of imatinib and/or levodopa. Values are expressed as means ± SEM ( n = 4-10). # P < 0.05, ## P < 0.01 versus MPTP-treated mice. One-way ANOVA [ F DARPP-32-pThr75(3,29) = 5.529, F DARPP-32-pThr34(3,16) = 1.257, F DARPP-32(3,16) = 2.886] followed by the Scheffe post hoc test. Levodopa (5), levodopa (5 mg/kg); IMB (10), imatinib mesylate (10 mg/kg). (G) Western-blot analysis of striatal levels of c-Abl-pTyr412, and c-Abl in MPTP-treated mice 30 min after a single i.p. injection of imatinib and/or levodopa. Values are expressed as means ± SEM ( n = 8–11). # P < 0.05 versus MPTP-treated mice; One-way ANOVA [ F c-Abl-pTyr412(3,34) = 5.820, F c-Abl(3,29) = 0.240] followed by Scheffe post hoc test. Levodopa (5), levodopa (5 mg/kg); IMB (10), imatinib mesylate (10 mg/kg).

Article Snippet: Mice received a single i.p. injection of imatinib mesylate (10 or 25 mg/kg; LKT Laboratories, St. Paul, MN, United States) dissolved in 0.9% saline containing 10% dimethyl sulfoxide 3 days after the administration of MPTP or saline.

Techniques: Injection, Clinical Proteomics, Western Blot

Journal: Frontiers in Pharmacology

Article Title: c-Abl Inhibition Exerts Symptomatic Antiparkinsonian Effects Through a Striatal Postsynaptic Mechanism

doi: 10.3389/fphar.2018.01311

Figure Lengend Snippet: Effects of imatinib on striatal presynaptic dopaminergic markers in MPTP-treated mice. Western-blot and HPLC analyses were carried out on the striatal extracts from vehicle or MPTP-treated mice 30 min after a single i.p. injection of imatinib mesylate (10 or 25 mg/kg). (A) Western-blot analysis of striatal levels of TH. Values are means ± SEM ( n = 4-5). ∗ P < 0.05 versus vehicle-treated mice; one-way ANOVA [ F (4,19) = 107.43] followed by the Scheffe post hoc test. IMB (10), imatinib mesylate (10 mg/kg); IMB (25), imatinib mesylate (25 mg/kg). (B) Western-blot analysis of striatal levels of DAT. Values are means ± SEM ( n = 4–5). ∗ P < 0.05 versus vehicle-treated mice; one-way ANOVA [ F (4,19) = 21.749] followed by the Scheffe post hoc test. IMB (10), imatinib mesylate (10 mg/kg); IMB (25), imatinib mesylate (25 mg/kg). (C) Western-blot analysis of striatal levels of VMAT2. Values are means ± SEM ( n = 4–5). ∗ P < 0.05 versus vehicle-treated mice; one-way ANOVA [ F (4,19) = 20.615] followed by the Scheffe post hoc test. IMB (10), imatinib mesylate (10 mg/kg); IMB (25), imatinib mesylate (25 mg/kg). (D–G) HPLC analysis of striatal levels of DA (D) , DOPAC (E) , HVA (F) , and DA-turnover, which represents a net dopamine usage in striatum with (DOPAC + HVA)/DA (G) . Values are expressed as means ± SEM ( n = 4–5). ∗ P < 0.05 versus vehicle-treated mice; one-way ANOVA [ F DA(4,19) = 34.526, F DOPAC(4,19) = 15.383, F HV A(4,19) = 16.078, F DA-turnover(4,19) = 10.355] followed by the Scheffe post hoc test. IMB (10), imatinib mesylate (10 mg/kg); IMB (25), imatinib mesylate (25 mg/kg).

Article Snippet: Mice received a single i.p. injection of imatinib mesylate (10 or 25 mg/kg; LKT Laboratories, St. Paul, MN, United States) dissolved in 0.9% saline containing 10% dimethyl sulfoxide 3 days after the administration of MPTP or saline.

Techniques: Western Blot, Injection

Journal: bioRxiv

Article Title: Targeting the translational machinery in gastrointestinal stromal tumors (GIST) – a new therapeutic vulnerability

doi: 10.1101/2021.09.01.458633

Figure Lengend Snippet: Homoharringtonine inhibits nascent protein synthesis and reduces KIT protein expression in GIST cells. (A) Nascent protein synthesis of GIST cells treated with homoharringtonine (HHT, 0.1 μM; green), cycloheximide (CHX, 100 μg/ml; blue), KIT inhibitor (IM, 1μM for GIST882 and GIST-T1; sunitinib, SU, 1μM for GIST430 and GIST48; orange) or 0.1% DMSO control (red) for 1 h or 8 h. Cells were labelled with HPG during the last 30 min of drug treatment and incorporated cellular HPG linked to azide-modified Alexa488 was quantitated by flow cytometry. Unstained control cells are shown in black in the histogram. A change in nascent protein synthesis is indicated as a relative mean fluorescence value to the DMSO control in the bar graphs. Columns, mean + SE; *, p<0.05 in comparison to DMSO control; **, p<0.01 in comparison to control; ***, p<0.001 in comparison to control (Student’s t-test, 2-tailed). (B) Immunoblot analysis for KIT protein expression of imatinib (IM)-sensitive (GIST882, GIST-T1) and IM-resistant (GIST430, GIST48) GIST cells after treatment with the protein translation inhibitor cycloheximide (CHX; 30 μg/ml for 3 h). (C) Immunoblotting for KIT protein expression in IM-sensitive (GIST882, GIST-T1) and IM-resistant (GIST430, GIST48) GIST cells after treatment with HHT for 72 h at the indicated concentrations. Abbreviations: 882 (GIST882), T1 (GIST-T1), 430 (GIST430), 48 (GIST48). (B, C) Grouped immunoblot images are either cropped from different parts of the same gel or from a separate gel run with another aliquot of the same protein lysate

Article Snippet: Homoharringtonine (HHT; Santa Cruz) treatments were performed at the indicated concentrations (in DMSO) compared to 0.1% DMSO for up to 72 h. Treatment with imatinib mesylate (1 µM in DMSO; LC Laboratories) or sunitinib (1 µM in DMSO; LC Laboratories) served as control.

Techniques: Expressing, Modification, Flow Cytometry, Fluorescence, Western Blot