Figure 3. Nupr1 knockdown attenuated sevoflurane-induced apoptosis and decreased C/EBPβ and IGFBP5 expression in HCN-2 neuronal cells. HCN-2 neuronal cells were transfected with NC or Nupr1 siRNA and then exposed to fresh gas (21% O2, 5% CO2, remainder N2) alone or with the addition of 3.4% sevoflurane for 6 h. (A, B) Western blotting for Nupr1, C/EBPβ, and IGFBP5 in 3.4% SEVO-exposed vs. control HCN-2 cells; representative images (A) and quantification (B). (C, D) Western blotting for Cleaved-Caspase-3, Bax, and Bcl-2 in 3.4% SEVO-exposed vs. control HCN-2 cells; representative images (C) and quantification (D). (E, F) Annexin V-FITC/PI staining and flow cytometry analysis; representative images (E) and quantification (F). **p<0.05. N=6.