Journal: International Journal of Molecular Sciences

Article Title: Sensitivity of Pancreatic Cancer Cell Lines to Clinically Approved FAK Inhibitors: Enhanced Cytotoxicity Through Combination with Oncolytic Coxsackievirus B3

doi: 10.3390/ijms26146877

Figure Lengend Snippet: Detection of Tyr-397 phosphorylated FAK and total FAK levels in Beta-TC-3 cells following treatment with FAKi. Upper panels: Beta-TC-3 cells were treated with 10 µM or 30 µM of the indicated FAKi (VS-4718, Ifebemtinib, VS-6062, GSK2256098, CEP-37440, or Defactinib) or with DMSO (0.1% or 0.3%) as a vehicle control. After 24 h, protein lysates were analyzed by Western blotting for Tyr-397 phosphorylated FAK (Tyr397-p125FAK) and total FAK (p125FAK). β-actin was used as an internal loading control. Arrows indicate bands corresponding to the expected molecular weight of 125 kDa. Lower panels: Quantification of the indicated protein levels was performed by densitometric analysis using ImageJ version 1.53a software. Expression levels of phosphorylated Tyr-397 FAK and total FAK were normalized to β-actin and are shown relative to DMSO-treated control cells, which were set to 100%. Shown are mean values ± SD from 2 samples. Notable both phosphorylated and total FAK proteins were not only detected at the expected molecular weight of 125 kDa but also at higher and lower molecular weights, suggesting the presence of multiple FAK isoforms in Beta-TC-3 cells [ , ].

Article Snippet: The Inhibitors of the FAK VS-4718 (PND-1186), Ifebemtinib (BI-853520), VS-6062 (PF-00562271), GSK2256098, CEP-37440, and Defactinib (VS-6063) were purchased as 100% DMSO dissolved solution (10 mM) from MedChemExpress (Sollentuna, Sweden) and stored as aliquots at −80 °C up to use.

Techniques: Control, Western Blot, Molecular Weight, Software, Expressing

Journal: International Journal of Molecular Sciences

Article Title: Sensitivity of Pancreatic Cancer Cell Lines to Clinically Approved FAK Inhibitors: Enhanced Cytotoxicity Through Combination with Oncolytic Coxsackievirus B3

doi: 10.3390/ijms26146877

Figure Lengend Snippet: Cytotoxic activity of FAKi with dose–response curves in pancreatic tumor cells. The five pancreatic tumor cell lines MIA Paca-2, BxPC-3, Capan-1, AsPC-1, and Beta-TC-3 were incubated with FAKi VS-4718, Ifebemtinib, VS-6062, GSK2256098, CEP-37440, or Defactinib at concentrations of 0.1–500 µM. Cell viability was determined by XTT assay 72 h post-treatment. Data were set relative to DMSO-treated cells. Shown are the nonlinear fits of each FAKi with mean values ± SD from 3 independent experiments.

Article Snippet: The Inhibitors of the FAK VS-4718 (PND-1186), Ifebemtinib (BI-853520), VS-6062 (PF-00562271), GSK2256098, CEP-37440, and Defactinib (VS-6063) were purchased as 100% DMSO dissolved solution (10 mM) from MedChemExpress (Sollentuna, Sweden) and stored as aliquots at −80 °C up to use.

Techniques: Activity Assay, Incubation, XTT Assay

Journal: International Journal of Molecular Sciences

Article Title: Sensitivity of Pancreatic Cancer Cell Lines to Clinically Approved FAK Inhibitors: Enhanced Cytotoxicity Through Combination with Oncolytic Coxsackievirus B3

doi: 10.3390/ijms26146877

Figure Lengend Snippet: Influence of FAKi on the replication and cytotoxicity of PD-H. ( A ) Viral growth curves. MIA Paca-2 and Capan-1 cells were infected with 3 and 0.1 MOI of PD-H for 1 h and thereafter incubated with medium containing the FAKi VS-4718, VS-6062, CEP-37440, or Defactinib at indicated concentrations, or DMSO, or cell culture medium alone for 24 h. Virus titers were determined by plaque assay on HeLa cells. Shown are mean values ± SD from 3 independent experiments. Statistical significance compared to PD-H + DMSO, * p < 0.05, ** p < 0.01, and *** p < 0.001; n.s., not significant. ( B ) Cell viability of the PD-H/FAKi approach described under ( A ). Cell viability was determined by XTT assay 24 h post-treatment. Data were set relative to DMSO-treated cells. Statistical significance compared to non-treated cells, * p < 0.05 and *** p < 0.001; n.s., not significant.

Article Snippet: The Inhibitors of the FAK VS-4718 (PND-1186), Ifebemtinib (BI-853520), VS-6062 (PF-00562271), GSK2256098, CEP-37440, and Defactinib (VS-6063) were purchased as 100% DMSO dissolved solution (10 mM) from MedChemExpress (Sollentuna, Sweden) and stored as aliquots at −80 °C up to use.

Techniques: Infection, Incubation, Cell Culture, Virus, Plaque Assay, XTT Assay

Journal: Genetics

Article Title: More than meets the eye: mutation of the white gene in Drosophila has broad phenotypic and transcriptomic effects

doi: 10.1093/genetics/iyaf097

Figure Lengend Snippet: Mutation of the white gene affects triglyceride levels and starvation resistance in a sex-specific manner. a) Body weight per fly of w + and w − males and females. b) Triglyceride content per body weight of fly compared for males and females. For a) and b), N = 5 replicates of 10 pooled flies. c) Starvation resistance measured by the number of hours for all flies in a replicate to die. N = 5 replicates of 20 flies each. d) Lipid peroxidation measured by absorbance ratio using BODIPY 581/591 C11 (Invitrogen). N = 20 replicates of 10 individuals each. All error bars represent the mean with 95% confidence interval.

Article Snippet: Sixty-three-microliter 2× TSS, 60-μL BODIPY solution (1 μL of 5 μm BODIPY 581/591 C11 lipid peroxidation sensor in DMSO per 1-mL ddH 2 O), and approximately 10–20 Lysing Matrix D beads (MP Biomedicals) were added to each sample.

Techniques: Mutagenesis