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Downstream molecular mechanisms and phenotypic consequences of SRC ablation in pemetrexed‐resistant cell lines. (A) SRC abrogation through transient transfection with SRC‐targeted CRISPR/Cas9 plasmids in resistant cells showed dose‐dependent reduction in SRC and p‐SRC Y416. Decreased p‐STAT3 and p‐FAK, as well as increased γ‐H2AX and cleaved caspase‐3, were observed. (B) Evaluation of the reciprocal regulation between SRC and TS, showing TS expression levels altered by siSRC transfection. (C) Flow cytometry showed increased Annexin V‐positive apoptotic cells after doxycycline‐induced conditional SRC knockout in resistant cell lines. (D) CFA showed reduced clonogenic capacity in resistant cell lines following conditional SRC ablation.
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Downstream molecular mechanisms and phenotypic consequences of SRC ablation in pemetrexed‐resistant cell lines. (A) SRC abrogation through transient transfection with SRC‐targeted CRISPR/Cas9 plasmids in resistant cells showed dose‐dependent reduction in SRC and p‐SRC Y416. Decreased p‐STAT3 and p‐FAK, as well as increased γ‐H2AX and cleaved caspase‐3, were observed. (B) Evaluation of the reciprocal regulation between SRC and TS, showing TS expression levels altered by siSRC transfection. (C) Flow cytometry showed increased Annexin V‐positive apoptotic cells after doxycycline‐induced conditional SRC knockout in resistant cell lines. (D) CFA showed reduced clonogenic capacity in resistant cell lines following conditional SRC ablation.
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Downstream molecular mechanisms and phenotypic consequences of SRC ablation in pemetrexed‐resistant cell lines. (A) SRC abrogation through transient transfection with SRC‐targeted CRISPR/Cas9 plasmids in resistant cells showed dose‐dependent reduction in SRC and p‐SRC Y416. Decreased p‐STAT3 and p‐FAK, as well as increased γ‐H2AX and cleaved caspase‐3, were observed. (B) Evaluation of the reciprocal regulation between SRC and TS, showing TS expression levels altered by siSRC transfection. (C) Flow cytometry showed increased Annexin V‐positive apoptotic cells after doxycycline‐induced conditional SRC knockout in resistant cell lines. (D) CFA showed reduced clonogenic capacity in resistant cell lines following conditional SRC ablation.
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Downstream molecular mechanisms and phenotypic consequences of SRC ablation in pemetrexed‐resistant cell lines. (A) SRC abrogation through transient transfection with SRC‐targeted CRISPR/Cas9 plasmids in resistant cells showed dose‐dependent reduction in SRC and p‐SRC Y416. Decreased p‐STAT3 and p‐FAK, as well as increased γ‐H2AX and cleaved caspase‐3, were observed. (B) Evaluation of the reciprocal regulation between SRC and TS, showing TS expression levels altered by siSRC transfection. (C) Flow cytometry showed increased Annexin V‐positive apoptotic cells after doxycycline‐induced conditional SRC knockout in resistant cell lines. (D) CFA showed reduced clonogenic capacity in resistant cell lines following conditional SRC ablation.
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Downstream molecular mechanisms and phenotypic consequences of SRC ablation in pemetrexed‐resistant cell lines. (A) SRC abrogation through transient transfection with SRC‐targeted CRISPR/Cas9 plasmids in resistant cells showed dose‐dependent reduction in SRC and p‐SRC Y416. Decreased p‐STAT3 and p‐FAK, as well as increased γ‐H2AX and cleaved caspase‐3, were observed. (B) Evaluation of the reciprocal regulation between SRC and TS, showing TS expression levels altered by siSRC transfection. (C) Flow cytometry showed increased Annexin V‐positive apoptotic cells after doxycycline‐induced conditional SRC knockout in resistant cell lines. (D) CFA showed reduced clonogenic capacity in resistant cell lines following conditional SRC ablation.
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Downstream molecular mechanisms and phenotypic consequences of SRC ablation in pemetrexed‐resistant cell lines. (A) SRC abrogation through transient transfection with SRC‐targeted CRISPR/Cas9 plasmids in resistant cells showed dose‐dependent reduction in SRC and p‐SRC Y416. Decreased p‐STAT3 and p‐FAK, as well as increased γ‐H2AX and cleaved caspase‐3, were observed. (B) Evaluation of the reciprocal regulation between SRC and TS, showing TS expression levels altered by siSRC transfection. (C) Flow cytometry showed increased Annexin V‐positive apoptotic cells after doxycycline‐induced conditional SRC knockout in resistant cell lines. (D) CFA showed reduced clonogenic capacity in resistant cell lines following conditional SRC ablation.

Journal: Thoracic Cancer

Article Title: SRC Activation Drives Acquired Pemetrexed Resistance in Non‐Small Cell Lung Cancer and Is Targetable by Dasatinib

doi: 10.1111/1759-7714.70406

Figure Lengend Snippet: Downstream molecular mechanisms and phenotypic consequences of SRC ablation in pemetrexed‐resistant cell lines. (A) SRC abrogation through transient transfection with SRC‐targeted CRISPR/Cas9 plasmids in resistant cells showed dose‐dependent reduction in SRC and p‐SRC Y416. Decreased p‐STAT3 and p‐FAK, as well as increased γ‐H2AX and cleaved caspase‐3, were observed. (B) Evaluation of the reciprocal regulation between SRC and TS, showing TS expression levels altered by siSRC transfection. (C) Flow cytometry showed increased Annexin V‐positive apoptotic cells after doxycycline‐induced conditional SRC knockout in resistant cell lines. (D) CFA showed reduced clonogenic capacity in resistant cell lines following conditional SRC ablation.

Article Snippet: BD Biosciences (San Diego, CA, USA) supplied fluorescein isothiocyanate (FITC)‐conjugated annexin V, and Selleckchem (Houston, TX, USA) supplied bosutinib, dasatinib, pemetrexed, and saracatinib.

Techniques: Transfection, CRISPR, Expressing, Flow Cytometry, Knock-Out