pwzl hygro fak y925f  (Addgene inc)

Journal: EMBO Molecular Medicine

Article Title: ERK5 suppression overcomes FAK inhibitor resistance in mutant KRAS-driven non-small cell lung cancer

doi: 10.1038/s44321-024-00138-7

Figure Lengend Snippet: Reagents and tools table

Article Snippet: pWZL-hygro-FAK Y925F , This study , Deposited at Addgene, plasmid #216547.

Techniques: Recombinant, Plasmid Preparation, Sequencing, shRNA, Reverse Transcription Polymerase Chain Reaction, Transfection, Western Blot, In Vitro, In Vivo, cDNA Synthesis, TUNEL Assay, Software

Journal: EMBO Molecular Medicine

Article Title: ERK5 suppression overcomes FAK inhibitor resistance in mutant KRAS-driven non-small cell lung cancer

doi: 10.1038/s44321-024-00138-7

Figure Lengend Snippet: Reagents and tools table

Article Snippet: pWZL-hygro-FAK Y861F , This study , Deposited at Addgene, plasmid #216546.

Techniques: Recombinant, Plasmid Preparation, Sequencing, shRNA, Reverse Transcription Polymerase Chain Reaction, Transfection, Western Blot, In Vitro, In Vivo, cDNA Synthesis, TUNEL Assay, Software

Journal: EMBO Molecular Medicine

Article Title: ERK5 suppression overcomes FAK inhibitor resistance in mutant KRAS-driven non-small cell lung cancer

doi: 10.1038/s44321-024-00138-7

Figure Lengend Snippet: Reagents and tools table

Article Snippet: pWZL-hygro-FAK S722A , This study , Deposited at Addgene, plasmid #216545.

Techniques: Recombinant, Plasmid Preparation, Sequencing, shRNA, Reverse Transcription Polymerase Chain Reaction, Transfection, Western Blot, In Vitro, In Vivo, cDNA Synthesis, TUNEL Assay, Software

Journal: EMBO Molecular Medicine

Article Title: ERK5 suppression overcomes FAK inhibitor resistance in mutant KRAS-driven non-small cell lung cancer

doi: 10.1038/s44321-024-00138-7

Figure Lengend Snippet: ( A ) Scheme depicting the critical dependency of KRAS mutant cancer cells on FAK. ( B ) Scheme showing the stepwise preparation of the HBEC3 cellular model. ( C ) Immunoblot against FAK in HBEC3 parental, HBEC3 mixed population before GFP + cell FACS sorting and HBEC3 FAK knockout clone. ( D ) Immunoblot analysis in HBEC3-FAK KO cell line transduced either with empty vector (pWZL-Hygro) or wild-type FAK (pWZL-Hygro-FAK WT) in the absence or presence of mutKRAS G12D (pBABE-zeo or pBABE-zeo-KRAS G12D ); mutKRAS: mutant KRAS. ( E ) Relative cell number of HBEC3-FAK KO cell line transduced as in ( D ); n = 3. ( F ) Relative quantification of cell death by flow cytometry analysis of Annexin V-Atto 633 (AV) -positive + Annexin V/PI (AV/PI)-positive + PI-positive gated populations in HBEC3-FAK KO cells previously transduced as in ( D ); n = 4. ( G ) Relative cell number of HBEC3-FAK KO cell line transduced either with wild-type FAK (pWZL Hygro-FAK WT) or FAK S732A (pWZL-Hygro-FAK S732A ) in the absence or presence of mutant KRAS G12D (pBABE-zeo or pBABE-zeo-KRAS G12D ). Mutant KRAS: mutKRAS; n = 3. ( H ) Relative cell number of HBEC3-FAK KO cell line transduced either with wild-type FAK (pWZL Hygro-FAK WT) or FAK S910A (pWZL-Hygro-FAK S910A ) in the absence or presence of mutKRAS G12D (pBABE-zeo or pBABE-zeo-KRAS G12D ). Mutant KRAS: mutKRAS; n = 3. ( I ) Relative quantification of colony forming capacity of HBEC3-FAK KO cells previously transduced as indicated; n = 3. ( J ) Quantification of cell death by flow cytometry analysis of Annexin V-Atto 633 (AV)-positive + Annexin V/PI (AV/PI)-positive + PI-positive HBEC3-FAK KO cells previously transduced as indicated; n = 3. Graphical data are mean ± SD. Statistical analyses were done using one-way ANOVA; n , number of biologically independent samples. .

Article Snippet: pWZL-hygro-FAK S732A , This study , Deposited at Addgene, plasmid #216541.

Techniques: Mutagenesis, Western Blot, Knock-Out, Plasmid Preparation, Quantitative Proteomics, Flow Cytometry

Journal: EMBO Molecular Medicine

Article Title: ERK5 suppression overcomes FAK inhibitor resistance in mutant KRAS-driven non-small cell lung cancer

doi: 10.1038/s44321-024-00138-7

Figure Lengend Snippet: Reagents and tools table

Article Snippet: pWZL-hygro-FAK S732A , This study , Deposited at Addgene, plasmid #216541.

Techniques: Recombinant, Plasmid Preparation, Sequencing, shRNA, Reverse Transcription Polymerase Chain Reaction, Transfection, Western Blot, In Vitro, In Vivo, cDNA Synthesis, TUNEL Assay, Software

Journal: EMBO Molecular Medicine

Article Title: ERK5 suppression overcomes FAK inhibitor resistance in mutant KRAS-driven non-small cell lung cancer

doi: 10.1038/s44321-024-00138-7

Figure Lengend Snippet: ( A ) Scheme depicting the critical dependency of KRAS mutant cancer cells on FAK. ( B ) Scheme showing the stepwise preparation of the HBEC3 cellular model. ( C ) Immunoblot against FAK in HBEC3 parental, HBEC3 mixed population before GFP + cell FACS sorting and HBEC3 FAK knockout clone. ( D ) Immunoblot analysis in HBEC3-FAK KO cell line transduced either with empty vector (pWZL-Hygro) or wild-type FAK (pWZL-Hygro-FAK WT) in the absence or presence of mutKRAS G12D (pBABE-zeo or pBABE-zeo-KRAS G12D ); mutKRAS: mutant KRAS. ( E ) Relative cell number of HBEC3-FAK KO cell line transduced as in ( D ); n = 3. ( F ) Relative quantification of cell death by flow cytometry analysis of Annexin V-Atto 633 (AV) -positive + Annexin V/PI (AV/PI)-positive + PI-positive gated populations in HBEC3-FAK KO cells previously transduced as in ( D ); n = 4. ( G ) Relative cell number of HBEC3-FAK KO cell line transduced either with wild-type FAK (pWZL Hygro-FAK WT) or FAK S732A (pWZL-Hygro-FAK S732A ) in the absence or presence of mutant KRAS G12D (pBABE-zeo or pBABE-zeo-KRAS G12D ). Mutant KRAS: mutKRAS; n = 3. ( H ) Relative cell number of HBEC3-FAK KO cell line transduced either with wild-type FAK (pWZL Hygro-FAK WT) or FAK S910A (pWZL-Hygro-FAK S910A ) in the absence or presence of mutKRAS G12D (pBABE-zeo or pBABE-zeo-KRAS G12D ). Mutant KRAS: mutKRAS; n = 3. ( I ) Relative quantification of colony forming capacity of HBEC3-FAK KO cells previously transduced as indicated; n = 3. ( J ) Quantification of cell death by flow cytometry analysis of Annexin V-Atto 633 (AV)-positive + Annexin V/PI (AV/PI)-positive + PI-positive HBEC3-FAK KO cells previously transduced as indicated; n = 3. Graphical data are mean ± SD. Statistical analyses were done using one-way ANOVA; n , number of biologically independent samples. .

Article Snippet: pWZL-hygro-FAK S910A , This study , Deposited at Addgene, plasmid #216542.

Techniques: Mutagenesis, Western Blot, Knock-Out, Plasmid Preparation, Quantitative Proteomics, Flow Cytometry

Journal: EMBO Molecular Medicine

Article Title: ERK5 suppression overcomes FAK inhibitor resistance in mutant KRAS-driven non-small cell lung cancer

doi: 10.1038/s44321-024-00138-7

Figure Lengend Snippet: Reagents and tools table

Article Snippet: pWZL-hygro-FAK S910A , This study , Deposited at Addgene, plasmid #216542.

Techniques: Recombinant, Plasmid Preparation, Sequencing, shRNA, Reverse Transcription Polymerase Chain Reaction, Transfection, Western Blot, In Vitro, In Vivo, cDNA Synthesis, TUNEL Assay, Software