Journal: bioRxiv
Article Title: Targeted KRAS G12V degradation elicits efficient and durable lung adenocarcinoma regression in vivo
doi: 10.1101/2024.12.13.627539
Figure Lengend Snippet: A, Colony forming assay in cells derived from dTAG-13 degradation-resistant LUADs. Paclitaxel = 1 µM (minimum dose to affect control cells). B, Degradation of dTAG- KRAS G12V in LUAD murine cells derived from mice treated with DMSO or the indicated compounds. (dTAG-13 = 0.3 µM, Tariquidar = 1 µM; 24h). ACTIN is shown as loading control. C, Degradation of dTAG-KRAS G12V in LUAD murine cells treated with DMSO or dTAG-13 (0.3 µM, 24h). Cell lines established from treatment-naïve (Controls) and dTAG-13 resistant LUAD tumors. Quantification in . D, Degradation of dTAG-KRAS G12V in LUAD murine cells treated with DMSO or dTAG V -1 (0.3 µM, 24h). Quantification in . E, DDB1 levels in cell lines established from treatment-naïve and dTAG-13 resistant LUAD tumors. ACTIN is shown as loading control. F, p38 degradation upon PROTAC treatment (1 µM; 16h) with NR-7h and NR-11c. Cell lines established from treatment-naïve and dTAG- 13 resistant LUAD tumors. ACTIN is shown as loading control. G, Frequency of alternative allele in SNPs or INDELs predicted to have a high or moderate impact in protein function. SNP calling with GATK Haplotype caller and annotation with SnpEff (see Methods). BL6: tumors from immunocompetent mice (C57BL/6J). NSG: tumors from immunocompromised mice. UT: untreated (red). T(sensitive): dTAG-13 sensitive tumors (green). T(resistant): dTAG-13 resistant tumors or derived cell lines (blue). BL6 UT tumors n = 4, sensitive n = 4, resistant n = 8. Cell lines UT n = 3, resistant n = 6. NSG UT tumors n = 4, sensitive n = 4, resistant n = 6. Only samples with variants with a minimum coverage of 10 reads are shown.
Article Snippet: The primary antibodies used are the following: CUL4A (#2699), DDB1 (#5428), NAE1 (#14321), CRBN (#71810) and p38 (#9217) were purchased from Cell Signaling Technology and used at 1:1000 dilution. β-ACTIN (A5441, used at 1:20000 dilution) and pan-RAS (OP40, used at 1:1000) were purchased from Sigma-Aldrich.
Techniques: Derivative Assay, Control
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