Fig. 9

TWEAK and TNF-α synergistically reduce the viability of auditory cells. (A) Experimental paradigm to assess the synergistic cytotoxic effects of TWEAK and TNF-α on auditory cells. (B) TWEAK and TNF-α co-treatment within the concentration range corresponding to their VS-secreted levels (TWEAK: 50 ng/mL; TNF-α: 0.05, 0.1, and 1 ng/mL) induced cell death in UB-OC1 and US-VOT-N33 cell lines, representing pro-sensory precursors of hair cells and neuroblasts, respectively. Cell death was using CellTox Green Dye staining with live cell imaging and expressed as a mean of fluorescence intensity relative to control-untreated cells. (C) TWEAK and TNF-α had a synergistic cytotoxic effect on auditory cells when exposed to equal concentrations of cytokines (50 ng/mL). Cell death is expressed as green integrated intensity per mm². *P < 0.05, **P < 0.01, ***P < 0.001, two-way ANOVA followed by Bonferroni’s multiple comparisons test (B), one-way ANOVA followed by Bonferroni’s multiple comparisons test (C). Each data point represents technical replicates; data are shown as mean ± s.e.m. GCU, green calibrated unit; VS, vestibular schwannoma