iAF cells have a differential ROS response to TNFα compared with NP cells.

(A) Representative images of CellROX green staining in NP cells exposed to TNFα (40 ng/mL). (B) Quantification of CellROX green average fluorescence intensity per cell in NP cells exposed to TNFα. (C) AmplexRed assay quantification of H2O2 released from NP cells exposed to TNFα. (D) Representative images of CellROX green staining in iAF cells exposed to TNFα (40 ng/mL). (E) Quantification of CellROX green average fluorescence intensity per cell in iAF cells exposed to TNFα. (F) AmplexRed quantification of H2O2 released from iAF cells exposed to TNFα. (G) Gene expression analysis relative to 18s rRNA of ROS related genes in NP cells exposed to TNFα. 18S rRNA Ct value (mean±SD): Control = 9.56± 0.5, TNFα = 9.55±0.36. (H) Gene expression analysis relative to 18s rRNA of ROS related genes in iAF cells exposed to TNFα. 18S rRNA Ct value (mean±SD): Control = 8.93±0.80, TNFα = 9.86±0.36. (I) AmplexRed quantification of H2O2 released from NP cells exposed to TNFα pre-treated with the NOX-inhibitor diphenyleneiodonium chloride (DPI). (J) AmplexRed quantification of H2O2 released from iAF cells exposed to TNFα pre-treated with the NOX-inhibitor DPI. Scale bars = 100μm. p<0.05 = *, p<0.01 = **, p<0.001 = ***, p<0.0001 = ****, N = 3 for all experiments.