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. 2024 May 22;15(5):358. doi: 10.1038/s41419-024-06730-6

Fig. 5. ET-1 facilitates heterotypic spheroid formation, survival, and invasive behavior.

Fig. 5

A Representative WB of whole cell lysates from HOFs, stimulated with ET-1 and/or AMB and/or BQ788 and/or BOS for 5 min, and probed with Abs to pMAPK, MAPK, pAKT and AKT. Histograms represent the mean ± SD of densitometric analyses of proteins relative to GAPDH; statistics were obtained using One-way ANOVA. B sh-SCR or sh-ARRB1 Kuramochi/HOFs treated with ET-1 and/or AMB and/or BQ788 and/or BOS were allowed to invade fibronectin/type I collagen plugs in an inverted invasion assay (48 h). Cells were stained with PKH67 (green, HOFs) or PKH26 (red, Kuramochi), before invasion assay, and serial optical sections (10 μm intervals) were acquired. The invasion was measured by dividing the sum of the signal intensity of all slides beyond 20 μm (invading cells) by the sum of the intensity of all slides (total cells); statistics were obtained using One-way ANOVA, Dunnett post hoc analysis. Scale bar, 100 µm.