Fig. 7

NAT10 activated the TNC/Akt/TGF-β1positive feedback loop to drive the malignant phenotypes of GC cells. a The protein expression of apoptosis-related protein markers in stable NAT10-overexpressing or NAT10-knockdown cell lines was measured by Western blotting analysis. NAT10 overexpression resulted in increased expression of Bcl2 and decreased expression of Bax, Cleaved Caspase-9 and Cleaved Caspase-3, which could be partly reversed by siTNC, while NAT10 knockdown caused the opposite alteration. b The protein expression of EMT-related protein markers in stable NAT10-overexpressing or NAT10-knockdown cell lines was measured by Western blotting analysis. NAT10 overexpression resulted in increased expression of N-cadherin and Vimentin and decreased expression of E-cadherin, which could be partly reversed by siTNC, while NAT10 knockdown caused the opposite alteration. c Representative images of immunofluorescence staining for the indicated markers. NAT10 overexpression led to increased expression of TNC, p-Akt and TGF-β1, while NAT10 knockdown caused the opposite alteration. Original magnification: 400×. d Stable NAT10-overexpressing cells were treated with siTNC or LY294002, and stable NAT10-knockdown cells were treated with TNC overexpression plasmid, followed by Western blotting analysis of the expression of the indicated markers. siTNC or LY294002 partly reversed the upregulated expression of TNC, p-Akt and TGF-β1 caused by NAT10 overexpression, while TNC overexpression plasmid partly reversed the downregulated expression of TNC, p-Akt and TGF-β1 caused by NAT10 knockdown. e MKN28 cells were treated with TGF-β1, and MKN45 cells were treated with P144, followed by Western blotting analysis of the expression of the indicated markers. TGF-β1 could upregulate the expression of TNC and p-Akt, while P144 caused the opposite alteration