Fig. 5
HO-1 inhibits NLRP1 expression through ATF4. a Expression of ATF4 and CHOP in primary neurons were determined by immunoblot after 6-h H2O2 treatment. Left panel: representative immunoblot image. Right panel: statistical analysis for the expression of ATF4 and CHOP. Un: untreated control. N = 4 per group. ***p < 0.001 compared with “Un” group. b mRNA levels of ATF4 and NLRP1 in primary neurons. Primary neurons were transfected with different doses of ATF4 siRNA 48 h before treatment with H2O2 for additional 6 h. N = 3 per group. *p < 0.05; **p < 0.01; ***p < 0.001 compared with “0” group. c Neurons were treated as in b, followed by detection of neuronal death with TUNEL after 12-h H2O2 treatment. N = 6 per group. *p < 0.05; ***p < 0.001 compared with “0” group. d Protein level of ATF4 in primary neurons. Primary neurons were transduced with A-HO-1 or transfected with ATF4 siRNA for 24 h before treatment with H2O2 for additional 6 h. Un: untreated control. C: cells under transduction or transfection condition without AAV and siRNA. Si-ATF4: siRNA for ATF4. This is a representative of three independent experiments. e Neurons were treated as in d. mRNA level of ATF4 was determined by q-RTPCR. N = 5 per group. **p < 0.01; ***p < 0.001