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. 2019 Oct 7;15(10):e1008439. doi: 10.1371/journal.pgen.1008439

Fig 4. TK1 knockdown affects Rho GTPase activity by reducing the activation of ribonucleotide reductase in LUAD cells.

Fig 4

(A) Experimental design for the glutathione S-transferase (GST) pull-down assay used to analyze active RhoA. (B) Active RhoA was measured in the indicated LUAD cell lines expressing TK1 shRNA or NS shRNA control using GST pull-down assays and immunoblot analysis. GST-RBD was used as a control in the pull-down assay, and total RhoA in whole-cell lysates was used as a loading control for immunoblot analysis. (C) GTP/GDP ratios were measured in the indicated LUAD cell lines expressing TK1 shRNA or NS shRNA control using high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS). (D) Active RhoA was measured in the indicated LUAD cell lines expressing TK1 shRNA or NS shRNA using actin (green)/vinculin (red) immunofluorescence and confocal microscopy. Representative images are shown. Scale bar, 20 μm.