Journal: iScience

Article Title: Tracing specificity of immune landscape remodeling associated with distinct anticancer treatments

doi: 10.1016/j.isci.2025.112071

Figure Lengend Snippet:

Article Snippet: Another cohort was treated for 10 or 20 days, every day by intraperitoneal injection with Trametinib (1 mg/kg, MEK inhibitor; TargetMol) plus ABT-737 (30 mg/kg, BCL-XL inhibitor; TargetMol), both diluted in Cremophore and NaCl 0.9%.

Techniques: Recombinant, Software

Journal: Journal of Nuclear Medicine

Article Title: Synthesis of the PET Tracer 124 I-Trametinib for MAPK/ERK Kinase Distribution and Resistance Monitoring

doi: 10.2967/jnumed.120.241901

Figure Lengend Snippet: Receptor tyrosine kinase (RTK) pathway containing RAS and BRAF. MEK is found in receptor tyrosine kinase pathway downstream of both KRAS and BRAF. Trametinib is MEK1/2 inhibitor and Food and Drug Administration–approved for BRAFV600E mutant cancers. By inhibiting MEK activity, trametinib can reduce hyperproliferative signaling from KRAS or BRAF mutants, though on-target toxicity is observed even with wild-type KRAS and BRAF cells.

Article Snippet: Imaging Studies Mice injected with 124 I-trametinib were imaged at 0, 1, 3, 6, 12, 24, and 96 h after injection using a Siemens Inveon PET/CT scanner.

Techniques: Mutagenesis, Activity Assay

Journal: Journal of Nuclear Medicine

Article Title: Synthesis of the PET Tracer 124 I-Trametinib for MAPK/ERK Kinase Distribution and Resistance Monitoring

doi: 10.2967/jnumed.120.241901

Figure Lengend Snippet: Synthesis and cell line specificity of radioiodinated trametinib. (A) Synthesis scheme started from trametinib into boronato-pinacol precursor readily available for iodination. Radiosynthesis and purification were complete within 1.5 h, with radiochemical yield of 70% for 124I-trametinib and specific activity greater than 100 GBq/μmol. (B) HPLC purification shows that most radioiodinated product is distinct from boronato-precursor and identical to cold trametinib. (C) Trametinib is type-III MEK allosteric inhibitor designed to reversibly inhibit adenosine triphosphate from phosphorylating MEK1/2. 124I-trametinib addition to cell lines expressing mutant Ras and or mutant Braf reveal nearly 10-fold range of uptake after 2 h, all of which is blocked with 1 μg of cold trametinib. Human KRAS mutant (G12V, G12C, or G12D) or BRAFV600E cell lines retained higher percentages of 124I-trametinib, with double KRAS BRAF mutant having highest uptake, differentiating KRAS and BRAF mutant tissue from normal. Murine cancer cell lines were all similarly avid for 124I-trametinib except Raw264.7, representing normal KRAS and BRAF and having lower uptake. DMSO = dimethyl sulfoxide; RCY = radiochemical yield; SA = specific activity; UV/Vis = ultraviolet/visible light.

Article Snippet: Imaging Studies Mice injected with 124 I-trametinib were imaged at 0, 1, 3, 6, 12, 24, and 96 h after injection using a Siemens Inveon PET/CT scanner.

Techniques: Purification, Activity Assay, Expressing, Mutagenesis

Journal: Journal of Nuclear Medicine

Article Title: Synthesis of the PET Tracer 124 I-Trametinib for MAPK/ERK Kinase Distribution and Resistance Monitoring

doi: 10.2967/jnumed.120.241901

Figure Lengend Snippet: In vivo distribution of 124I-trametinib in healthy mice primarily in liver, with subsequent elimination through gastrointestinal tract. (A) PET imaging at 1, 3, 6, and 24 h shows broad distribution in tissues, with highest uptake in liver, spleen, and kidneys and later gastrointestinal clearance. Scale bar values in injected dose per gram (%ID/g). (B) Terminal biodistributions at up to 12 h show high heart and lung uptake (5 %ID/g) compared with blood (1 %ID/g) or skin (1–2 %ID/g). Peak uptake in skin was observed at 6 h after injection. GI = gastrointestinal.

Article Snippet: Imaging Studies Mice injected with 124 I-trametinib were imaged at 0, 1, 3, 6, 12, 24, and 96 h after injection using a Siemens Inveon PET/CT scanner.

Techniques: In Vivo, Imaging, Injection

Journal: Journal of Nuclear Medicine

Article Title: Synthesis of the PET Tracer 124 I-Trametinib for MAPK/ERK Kinase Distribution and Resistance Monitoring

doi: 10.2967/jnumed.120.241901

Figure Lengend Snippet: Imaging and biodistribution of 124I-trametinib in mice bearing B16F10 melanomas through 72 h. (A) PET imaging of B16F10 melanoma–bearing mice shows slow tumor uptake through 72 h. Scale bar values in injected dose per gram (%ID/g). (B) Terminal biodistribution reveals maximal uptake of 124I-trametinib in B16F10 tumor footpad between 48 and 72 h after injection. Inset: ratio of %ID/g in tumor to skin, blood, muscle, and heart increased after 24 h, showing retention of 124I-trametinib in tumor relative to other tissues. Tumor uptake after 24 h would allow imaging of naïve tumors in addition to monitoring tumor resistance during conventional trametinib therapy.

Article Snippet: Imaging Studies Mice injected with 124 I-trametinib were imaged at 0, 1, 3, 6, 12, 24, and 96 h after injection using a Siemens Inveon PET/CT scanner.

Techniques: Imaging, Injection

Journal: Journal of Nuclear Medicine

Article Title: Synthesis of the PET Tracer 124 I-Trametinib for MAPK/ERK Kinase Distribution and Resistance Monitoring

doi: 10.2967/jnumed.120.241901

Figure Lengend Snippet: Trametinib therapy reduces uptake of 124I-trametinib systemically. Prior administration of 6 mg kg−1 trametinib intraperitoneally once per day for 3 d shows, by PET, reduced uptake of 124I-trametinib at 24 h (A) and 48 h (B) after administration. Scale bar values in injected dose per gram (%ID/g). (C) Terminal biodistribution on post–48-h PET scan confirms significant decreases in 124I-trametinib uptake in liver, spleen, kidneys, total gastrointestinal tract, bladder, and right-flank B16F10 tumor. Two-way ANOVA was used for biodistribution analysis of 4 mice receiving ×3 trametinib treatment and 5 naïve mice before 124I-trametinib imaging. ****P < 0.001. **P < 0.01. *P < 0.05. LN = lymph node; TGI = total gastrointestinal tract; QD = once daily.

Article Snippet: Imaging Studies Mice injected with 124 I-trametinib were imaged at 0, 1, 3, 6, 12, 24, and 96 h after injection using a Siemens Inveon PET/CT scanner.

Techniques: Injection, Imaging

Journal: bioRxiv

Article Title: SHP2 Inhibition Abrogates MEK inhibitor Resistance in Multiple Cancer Models

doi: 10.1101/307876

Figure Lengend Snippet: A-B , Response of MiaPaCa-2 (A) and Capan-2 (B) subcutaneous xenografts to treatment with SHP099 (75 mg per kg body weight, daily), trametinib (0.25 mg per kg body weight, daily) or both drugs (trametinib 0.25 mg/kg, daily; SHP099 75 mg/kg every other day). Waterfall plot shows response of each tumor after 3 weeks of treatment; n = 8-10 mice per group. (** P < 0.005; *** P < 0.0005, two-tailed Mann Whitney test). C-D , Effects on p-ERK levels in MiaPaCa-2 tumors following 3 weeks of drug treatment, as shown by p-ERK immunoblotting (C) and immunohistochemical analysis (D). E , Syngeneic mice with orthotopic injections of FC1242 (KPC) cells were treated with vehicle, SHP099 (75 mg per kg body weight, daily), trametinib (0.25 mg per kg body weight, daily) or both drugs (trametinib 0.25 mg/kg, daily; SHP099 75 mg/kg every other day), as depicted in the schematic, and tumor size was measured at day 14 (*** P < 0.0005, one-tailed Mann Whitney test) F , Effects on p-ERK activity in FC1242-derived tumors from E. G , Representative haematoxylin and eosin (H&E) stains of drug-treated tumors from E. H , Sizes of tumors from syngeneic mice (C57BL/6J) and athymic nude ( nu/nu ) mice injected orthotopically with the same number of FC1242 (KPC) cells and treated with SHP099 (75 mg per kg body weight, daily), trametinib (1.0 mg per kg body weight, daily) or both drugs (trametinib 1.0 mg/kg + SHP099 75 mg/kg every other day), as depicted in the schematic, measured at day 22 (** P < 0.005, *** P < 0.0005, one-tailed Mann Whitney test). I , Note larger effects seen in immune-competent mice (* P < 0.05, ** P < 0.001; two-sided t test). Numbers under blots indicate relative intensities, compared with untreated controls, quantified by LICOR.

Article Snippet: Selumetinib-AZD6244 (S1008), UO126 (S1102) and trametinib (GSK1120212-S2673) were purchased from Selleckchem.

Techniques: Two Tailed Test, MANN-WHITNEY, Western Blot, Immunohistochemical staining, One-tailed Test, Activity Assay, Derivative Assay, Injection

Journal: bioRxiv

Article Title: SHP2 Inhibition Abrogates MEK inhibitor Resistance in Multiple Cancer Models

doi: 10.1101/307876

Figure Lengend Snippet: A-C , TNBC (A) and HGSC (B) cell lines were treated with DMSO, SHP099, MEK-I (U0126 or AZD6244), or both (COMBO). PrestoBlue (A) and proliferation (B) assays were performed at one week. Colony formation (C) assay was performed at two weeks. Representative results from a minimum of three biological replicates are shown per condition: SHP099 10 μM, U0126 (10 μM) or AZD6244 (1 μM) as indicated, Combo=SHP099 10 μM + UO126 (10 μM)/AZD6244 1 μM (* P < 0.05, ** P < 0.001, *** P < 0.0001, two-sided t test). Red asterisk indicates synergistic interaction between the two drugs by BLISS independent analysis. D , GST-RBD pull down assay from TNBC and HGSC cell line lysates treated with DMSO, SHP099 10 μM, AZD6244 1 μM, or both for 48 h. Image is representative of at least two independent experiments. E , Immunoblots of lysates from TNBC and HSGS cell lines, treated as indicated. Image is representative of three independent experiments. F , ERK-dependent gene expression ( ETV1,4, 5 and FOSL-1 ), assessed by qRT-PCR, in TNBC and HGSC lines treated for 48h with the indicated drugs. G-H , MDA-MB-468 (G) and PDX-2555 (B) mammary fat pad xenografts following treatment with SHP099 (75 mg per kg body weight, daily), trametinib (0.25 mg per kg body weight, daily) or both drugs (trametinib 0.25 mg/kg, daily; SHP099 75 mg/kg every other day). Waterfall representations of tumor responses after 3 weeks of treatment are shown; n = 8-10 mice per group. (** P < 0.005, *** P < 0.0005, two-tailed Mann Whitney test). Numbers under blots indicate relative intensities, compared with untreated controls, quantified by LICOR.

Article Snippet: Selumetinib-AZD6244 (S1008), UO126 (S1102) and trametinib (GSK1120212-S2673) were purchased from Selleckchem.

Techniques: Pull Down Assay, Western Blot, Gene Expression, Quantitative RT-PCR, Two Tailed Test, MANN-WHITNEY

Journal: Nature medicine

Article Title: Genomic profiling informs diagnoses and treatment in vascular anomalies

doi: 10.1038/s41591-023-02364-x

Figure Lengend Snippet: Transduction of BRAF-F486S (a) or RAF1-T145P (c) significantly increased the level of p-ERKs and Trametinib treatment led to a significant reduction of p-ERKs. Three-dimensional lymphatic spheroid sprouting assay showed elevated sprouting activity in HDLECs expressing BRAF-F486S (b) or RAF1-T145P (d) compared to its WT as measured by both cumulative sprout length and number of sprouts. Trametinib treatment led to a significant reduction of both cumulative sprout length and number of sprouts. For a-d, three independent experiments were performed. P values were calculated using 2-sided t-tests and included in each panel (degree of freedom was included in the Source Data for Extended Data Fig. 1), and corrected for multiple testing using the FDR (Benjamini and Hochberg) method. Bar graphs in panels a and c represent mean fold change in the ratio of pERK to GAPDH, normalized to the untreated wild type. Error bars represent standard deviations. In the box and whisker plots in panels b and d, the line in the middle of the box represents the medians, tops and bottoms of the boxes represent the 25th and 75th quartiles respectively, and the whiskers extend to 1.5 times the interquartile range beyond the 25th and 75th quartiles. All the data points that are summarized by the boxplots are superimposed as dots on the plots; minima and maxima can be determined by the highest and lowest dots. e, e’, Expression of KRASWT had no impact on lymphatic vessel morphology in trunk. f, f’, Expression of KRASA146T resulted in lymphatic tissue expansion (asterisks) and dilation of the thoracic duct (dotted line). Red: Expression of transgene in trunk of zebrafish at 5dpf, Green: mrc1a:GFF labeling lymphatic vessels. g, Quantitation of WT EK larvae that were assayed for pericardial edema at 5 dpf. Injected embryos were screened for transgenic mCherry expression in endothelial cells prior to quantitation so that only transgenic expressing embryos were counted. The p-values in the graph were calculated via Fisher’s Exact tests (two-sided) between samples as indicated, followed by Bonferroni correction. Indicated n are the total number of larvae assayed per condition.

Article Snippet: Primary human dermal lymphatic endothelial cells acquired from Promocell (catalog number C-12216; from juvenile foreskin) were retrovirally transduced with wild type or different variants and cultured for 48–72 h. Transduced cells were plated in a 96-well plate (20,000 cells per well) in the presence of carrier (0.1% DMSO) or 300 nM trametinib and cultured for 24 h. Cells were lysed, lysates were cleared by centrifugation and analyzed by SDS–PAGE and western blotting with pERK (Cell Signaling Technology no. 4376) and GAPDH (Santa Cruz Biotechnology no. sc-47724).

Techniques: Transduction, Activity Assay, Expressing, Whisker Assay, Labeling, Quantitation Assay, Injection, Transgenic Assay

Journal: Nature medicine

Article Title: Genomic profiling informs diagnoses and treatment in vascular anomalies

doi: 10.1038/s41591-023-02364-x

Figure Lengend Snippet: a, Of the participants in the study, 44% (69) were already on medical therapy or planned to initiate medical therapy, and 56% (87) were not on medical therapy. b, Distribution of the impact of the molecular finding on medical therapy. The molecular finding supported the chosen therapy in 25 participants and did not support the medication in 1 participant. The molecular diagnosis led to a change in therapy in 13 participants and 4 additional participants planned to change therapy based on the finding. Twelve participants commenced therapy based on the genetic diagnosis and 14 more are planning to initiate therapy. c, Response to therapy. Three participants had worsening of disease, 9 participants had stable disease, 35 participants had improvement in disease and 3 participants had a mixed response to disease. For five participants, it was too early to assess disease response or information was not available (not applicable, NA). d, D-dimer decreased from 37 mg L−1 FEU at the start (off graph) to below 5 mg L−1 FEU within two cycles (CVA14). e, Pulmonary function tests show an increase in forced vital capacity (FVC), forced residual capacity (FRC), forced expiratory volume (FEV1) and total lung capacity (TLC). f–h, MR images before and after trametinib treatment. DCMRL (f) and T2 space (g) demonstrate significant mediastinal perfusion (f, thin arrow) and thickening (g, thin arrow) as well as pleural effusions (g, thick arrow). These findings are significantly reduced post treatment (h, arrows).

Article Snippet: Primary human dermal lymphatic endothelial cells acquired from Promocell (catalog number C-12216; from juvenile foreskin) were retrovirally transduced with wild type or different variants and cultured for 48–72 h. Transduced cells were plated in a 96-well plate (20,000 cells per well) in the presence of carrier (0.1% DMSO) or 300 nM trametinib and cultured for 24 h. Cells were lysed, lysates were cleared by centrifugation and analyzed by SDS–PAGE and western blotting with pERK (Cell Signaling Technology no. 4376) and GAPDH (Santa Cruz Biotechnology no. sc-47724).

Techniques: Biomarker Discovery

Journal: Cancers

Article Title: IGF1R/IR Mediates Resistance to BRAF and MEK Inhibitors in BRAF-Mutant Melanoma

doi: 10.3390/cancers13225863

Figure Lengend Snippet: Growth kinetics and characteristics of TDR cell lines. ( A ) Schematic of TDR cell generation. ( B ) WM115, WM983B, and A375 parental and TDR cells (1 × 10 4 cells/well) were seeded in 96-well plates and treated with increasing concentrations of dabrafenib (D) and trametinib (T) (0.1 nM–5 μ M) for 72 h. Cells were treated with MTT for 4 h and absorbance was read at 570 nm. Data reflect three independent experiments and are presented as mean ± SEM. ( C ) Dose response MTT proliferation assay for parental and TDR cell lines maintained in drug-free conditions for 21 days. Cells were plated for 24 h and treated with 72 h with increasing concentrations of dabrafenib (D) and trametinib (T). Cells were treated with MTT. Data reflected as mean ± SEM. ( D ) Immunoblots of WM983B, WM115, and A375 parental and TDR cells treated for 4 and 24 h with dabrafenib (D), trametinib (T), or combination of dabrafenib and trametinib (D + T). Whole cell lysates from the cell lines were analyzed by Western blot using indicated antibodies. Actin served as the loading control. ( E ) Immunoblots for TDR cells treated with increasing concentrations of SCH772984 (5–100 nM) for 24 h. Actin was used as the loading control.

Article Snippet: Parental and TDR cells treated with DMSO or a combination of dabrafenib and trametinib were lysed and 400 μg proteins were used for Proteome Profiler Human Phospho-RTK Array Kit (R&D Systems, cat#ARY001B(Minneapolis, MN, USA)) as per the manufacturer’s instructions.

Techniques: Proliferation Assay, Western Blot, Control

Journal: Cancers

Article Title: IGF1R/IR Mediates Resistance to BRAF and MEK Inhibitors in BRAF-Mutant Melanoma

doi: 10.3390/cancers13225863

Figure Lengend Snippet: Upregulation of IGF1R and IR kinase activity in BRAF-mutant melanoma cells resistant to BRAF and MEK inhibitors. ( A ) Parental and TDR cells (WM115, WM983B, and A375) either treated with DMSO or combination of dabrafenib (D) and trametinib (T) (WM115: D: 800 nM, T: 200 nM; WM983B: D: 2.4 µM, T: 500 nM; A375: D: 250 nM, T: 12.5 nM) for 24 h. The red boxes indicate the receptors with altered tyrosine kinase activity. Each RTK spotted in duplicate with positive controls in the corner. ( B ) The average signal density of spots normalized to parental DMSO group; p -values were computed using GraphPad Prism 7.0. * p < 0.05; ** p < 0.001 by two-tailed Student’s t -test. Data are presented as mean ± SEM. ( C ) Immunoblot analysis of parental and TDR cells treated with DMSO or combination of dabrafenib and trametinib (D + T) for 24 h. Whole cells were analyzed using Western blot with indicated antibodies. Actin served as the loading control.

Article Snippet: Parental and TDR cells treated with DMSO or a combination of dabrafenib and trametinib were lysed and 400 μg proteins were used for Proteome Profiler Human Phospho-RTK Array Kit (R&D Systems, cat#ARY001B(Minneapolis, MN, USA)) as per the manufacturer’s instructions.

Techniques: Activity Assay, Mutagenesis, Two Tailed Test, Western Blot, Control

Journal: Cancers

Article Title: IGF1R/IR Mediates Resistance to BRAF and MEK Inhibitors in BRAF-Mutant Melanoma

doi: 10.3390/cancers13225863

Figure Lengend Snippet: IGF1R and INSR are differentially expressed in TDR cells. ( A ) Heatmap of differentially expressed genes in parental cells, parental cells treated with combination of dabrafenib (D) and trametinib (T) (WM115: D: 800 nM, T: 200 nM; WM983B: D: 2.4 µM, T: 500 nM; A375: D: 250 nM, T: 12.5 nM), or TDR cells. ( B ) Quantification of Actin-normalized mRNA levels using real-time quantitative PCR for IGF1R, IGF1, IGF2, and INSR for parental cells treated with DMSO or dabrafenib + trametinib and TDR following 24 h of treatment. * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001 by two-tailed Student’s t -test. Data are presented as mean ± SEM. ( C ) Kaplan Meier survival curve of skin cutaneous melanoma patients (TCGA, PanCancer Atlas) with data stratified for BRAF mutation along with IGF1R expression z -score > 2 ( n = 8) and IGF1R expression z -score < 2 ( n = 179) (Left panel); and INSR expression z -score > 2 ( n = 8) and INSR expression z -score < 2 ( n = 181) (Right panel).

Article Snippet: Parental and TDR cells treated with DMSO or a combination of dabrafenib and trametinib were lysed and 400 μg proteins were used for Proteome Profiler Human Phospho-RTK Array Kit (R&D Systems, cat#ARY001B(Minneapolis, MN, USA)) as per the manufacturer’s instructions.

Techniques: Real-time Polymerase Chain Reaction, Two Tailed Test, Mutagenesis, Expressing

Journal: Cancers

Article Title: IGF1R/IR Mediates Resistance to BRAF and MEK Inhibitors in BRAF-Mutant Melanoma

doi: 10.3390/cancers13225863

Figure Lengend Snippet: BMS-754807 in combination with dabrafenib and trametinib inhibits cell proliferation and inhibits intracellular Akt ( A ) TDR cells (WM115, WM983B, and A375) plated for 24 h and treated with DMSO, dabrafenib (D), and trametinib (T) (WM115: D: 800 nM, T: 200 nM; WM983B: D: 2.4 µM, T: 500 nM; A375: D: 250 nM, T: 12.5 nM) treated alone or in combination with BMS-754807 (3, 5, and 10 µM) and treated with MTT. The intensities are represented as mean. Error bars: SEM *** p < 0.001; **** p < 0.0001 by two-tailed Student’s t -test compared to DMSO group. ( B ) TDR cells were plated and treated with media containing DMSO, dabrafenib (D) and tramatenib (T) (WM115: D: 800 nM, T: 200 nM; WM983B: D: 2.4 µM, T: 500 nM; A375: D: 250 nM, T: 12.5 nM), or BMS-754807 (5 µM) in combination with dabrafenib and trametinib (D + T + BMS). Media containing drugs were replenished every second day and stained with 0.5% crystal violet on Day 21. Representative images of the well for each treatment (Top panel). Quantification of intensities (Bottom panel) for the wells is represented as mean. Error bars: SEM ( n = 2 independent experiments performed in triplicates). *** p < 0.001 by two-tailed Student’s t -test compared to DMSO group. ( C ) TDR cells were seeded on matrigel basement membrane and treated with indicated drugs every second day. Pictures of each well were captured on day 21 (left panel) at 10× magnification. Quantification of mean area of acini (right panel). **** p < 0.0001 by two-tailed Student’s t -test compared to DMSO group. ( D ) TDR cells were serum starved for 24 h and treated with BMS-754807 (5 µM) along with dabrafenib (D) and trametinib (T) (WM115 TDR: D: 800 nM, T: 200 nM; WM983B TDR: D: 2.4 µM, T: 500 nM; A375 TDR: D: 250 nM, T: 12.5 nM) for 4 or 24 h followed by IGF2 (10 nM) stimulation for 1 h. Whole cell lysates were analyzed by Western blot using indicated antibodies. Actin was used as loading control.

Article Snippet: Parental and TDR cells treated with DMSO or a combination of dabrafenib and trametinib were lysed and 400 μg proteins were used for Proteome Profiler Human Phospho-RTK Array Kit (R&D Systems, cat#ARY001B(Minneapolis, MN, USA)) as per the manufacturer’s instructions.

Techniques: Two Tailed Test, Staining, Membrane, Western Blot, Control

Journal: Cancers

Article Title: IGF1R/IR Mediates Resistance to BRAF and MEK Inhibitors in BRAF-Mutant Melanoma

doi: 10.3390/cancers13225863

Figure Lengend Snippet: BMS-754807 inhibits tumor growth in WM115 TDR xenograft model. ( A ) Image of WM115 parental and TDR xenograft injection in nude mice. ( B ) Tumor growth kinetics of WM115 parental ( n = 8) and TDR xenograft ( n = 8) for 20 days post tumor injection. ( C ) Nude mice were injected with WM115 TDR cells and treated with vehicle, dabrafenib + trametinib (D + T), BMS-754807 (BMS), or the combination (BMS + D + T). Treatment was administered for 2 weeks by oral route. Tumors were measured thrice a week with calipers. Each data point represents the mean tumor volume ± SEM ( n = 6–7). All data plotted as mean ± SEM. * p < 0.05, ** p < 0.005 calculated using Student’s t -test. ( D ) Mice weight was recorded post treatment with BMS-754807 in combination with dabrafenib and trametinib and represented. ( E ) p-IGF1R/p-IR and p-Akt expressions were assessed in the tumor lysate by Western blot. Actin served as loading control.

Article Snippet: Parental and TDR cells treated with DMSO or a combination of dabrafenib and trametinib were lysed and 400 μg proteins were used for Proteome Profiler Human Phospho-RTK Array Kit (R&D Systems, cat#ARY001B(Minneapolis, MN, USA)) as per the manufacturer’s instructions.

Techniques: Injection, Western Blot, Control

Journal: Developmental cell

Article Title: Sphingosine 1-phosphate receptor signaling establishes AP-1 gradients to allow for retinal endothelial cell specialization

doi: 10.1016/j.devcel.2020.01.016

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: For MEK inhibition, P6 neonates were subcutaneously injected with 1 mg/kg of Trametinib (GSK1120212, AdooQ Bioscience, CAT#A11029, 0.1 mg/mL solution) or vehicle (1% DMSO in corn oil).

Techniques: Purification, Control, Recombinant, Agarose Gel Electrophoresis, SYBR Green Assay, Staining, Imaging, Sequencing, RNA Sequencing, Software, Microscopy, Real-time Polymerase Chain Reaction, Injection

Journal: Developmental cell

Article Title: Sphingosine 1-phosphate receptor signaling establishes AP-1 gradients to allow for retinal endothelial cell specialization

doi: 10.1016/j.devcel.2020.01.016

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: For pharmacological inhibition of S1PR1, neonates were intragastrically injected with 25 mg/kg of Trametinib NIBR-0213 (Aobious, CAT#AOB1113) or vehicle (DMSO).

Techniques: Purification, Control, Recombinant, Agarose Gel Electrophoresis, SYBR Green Assay, Staining, Imaging, Sequencing, RNA Sequencing, Software, Microscopy, Real-time Polymerase Chain Reaction, Injection