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. 2016 Feb 15;6:20992. doi: 10.1038/srep20992

Figure 4. COX-2 signaling in RAW 264.7 cells in response to apoptotic cells mediates EMT inhibition in LA-4 cells.

Figure 4

(a,b,e) RAW 264.7 cells were pretreated with 10 μM NS-398 or 10 μM PD-146176 for 1 h and then stimulated with apoptotic Jurkat cells (ApoJ) for 20 h. (c) RAW cells were transfected with COX-2 siRNA or control vehicle (siRNA-GFP) for 6 h, then incubated with ApoJ for 20 h. Conditioned medium (CM) was added to LA-4 cells in the presence of TGF-β1 for 72 h. (a) Morphological changes in the cells were examined by phase-contrast microscopy. Scale bars = 50 μm. (b,c) Immunoblots of total cell lysates were performed with anti-E-cadherin, N-cadherin, or α-SMA antibodies. Right: Densitometric analysis of the indicated EMT markers’ relative abundances. (d) The amounts of the Snail1/2, Zeb1/2, and Twist1 mRNAs in LA-4 cell samples were analyzed by real-time PCR and normalized to that of Hprt mRNA. Values represent the mean ± s.e.m. of three independent experiments. *P < 0.05 compared with control; +P < 0.05 as indicated.