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. 2019 Mar 28;14(3):e0214553. doi: 10.1371/journal.pone.0214553

Fig 5. YAPS127A fibroblasts rearrange ECM structures and promote motility and invasiveness of OSCC cells.

Fig 5

(A) Fluorescence microscope was used to check the expression of EGFP on cultured cell after transfection with pcDNA4-HisMaxB-YAP1 and YAPS127A, Scale bar, 200μm. (B) The YAP, p-YAP(S127) and αSMA protein expression was identified in all transfected cells (empty-, WT-YAP-, YAPS127A transfected hTERT-hNOFs) by Western blot. (C) Immunofluorescence images were shown in WT-YAP1- and YAPS127A-hTERT-hNOFs. Merged stained Images (DAPI(blue), F-actin(Phalloidin)(red), YAP(green))are shown. Scale bar, 50μm. The bar graph indicates the distribution of YAP in WT-YAP1- and YAPS127A-hTERT-hNOFs. It was analyzed by Image J software program (WT-YAP1-hTERT-hNOFs, n = 40; YAPS127A-hTERT-hNOFs, n = 40) (***p < 0.001; Student t test). (D) The representative images show gel contractility by the fibroblasts. Histogram shows mean value ± SD (n = number of gels (Empty, 3; WT-YAP, 3; YAPS127A, 3), assessed over multiple experiments (*p < 0.05; Mann Whitney U test). (E) Histogram shows elastic modulus of matrix remodeled by fibroblasts, respectively. Each data point represents a single measurement, (n = 3, respectively) measurements in total. Line and error bars indicate mean value ± SD. (F-G) Representative images showed the (F) migration and (G) invasion of OSCC cells (YD10B and YD38) cultured with each of fibroblasts, which were transfected with Empty-, WT-YAP-, YAPS127A-transfected hTERT-hNOFs). Quantification of OSCC cells (YD10B and YD38) migration toward lower chamber is shown as bar graphs. Each assay was performed in independent triplicate repeats (*p < 0.05; Mann-Whitney U test).