Impaired inflammasome activation and bacterial clearance in G6PD deficiency due to defective NOX/p38 MAPK/AP-1 redox signaling

Yen, Wei-Chen; Wu, Yi-Hsuan; Wu, Chih-Ching; Lin, Hsin-Ru; Stern, Arnold; Chen, Shih-Hsiang; Shu, Jwu-Ching; Tsun-Yee Chiu, Daniel

The total and phosphorylated form of c-Fos, but not c-Jun, decreased with or without LPS treatment in G6PD-kd THP-1 cells (Fig. 5A) . The quantitative intensities of p-c-Fos and c-Fos are shown in Fig. 5B and C, respectively. AP-1 signaling, which is the heterodimer of c-Fos and c-Jun and is downstream of p38, decreased after treatment with SB203580 in both control and G6PD-kd THP-1 cells (Fig. 5D) . The quantitative intensity of c-Fos is shown in Fig. 5E . LPS treatment promoted the translocation of c-Jun, c-Fos and p65 from the cytoplasm to the nucleus in control and G6PD-kd THP-1 cells (Fig. 5F ). The translocation of c-Fos was reduced in G6PD-kd THP-1 cells, although c-Jun translocation was increased upon G6PD knockdown compared with that in the scrambled control cells. No significant difference was observed in the translocation of NF-κB. The quantification of nuclear c-Fos is shown in Fig. 5G . The IL1B reporter plasmid construction is shown in Fig. 5H , upper panel. 293FT cells were chosen to measure the binding activity of AP-1 because of the ease of transfection of these cells compared to that of THP-1 cells. The binding activity of AP-1 was lower in 293FT pLKO-G6PD-kd cells compared to the binding activity of 293FT pLKOscrambled cells (Fig. 5H) .