Figure 6

CUR regulates energy metabolism through AMPK-dependent pathways in PRV-infected BV2 cells. A BV2 cells were infected with/without PRV for 24 h and then treated with/without 20 μM CUR for 24 h. Western blot analysis of p-AMPK^Thr172 protein levels (normalized to the t-AMPK protein). β-Actin was used as the loading control (n = 3). B Relative protein levels of p-AMPK^Thr172. C BV2 cells were treated with different concentrations of Compound C for 24 h, and cell viability was determined by the CCK-8 assay to determine the toxicity range (n = 4). D BV2 cells were infected with/without PRV or were treated with PRV and an AMPK inhibitor (Compound C or siRNA) alone or in combination for 24 h, followed by treatment with/without 20 μM CUR for 24 h. Western blot analysis of p-AMPK (normalized to the t-AMPK protein), t-AMPK, Gpat4, and LDHa levels (Gpat4 and LDHa values were normalized to the β-actin protein); β-actin was used as the loading control (n = 3). E Relative p-AMPKThr172 protein levels. F Relative LDHa protein levels. G Relative Gpat4 protein levels. H The extracellular acidification rate of BV2 cells after the different treatments (values normalized to the control) (n = 4). I The OCR of BV2 cells after the different treatments (values normalized to the control) (n = 4). J BV2 cell ATP levels in the different treatment groups (n = 4). K The M1 phenotype-related inflammatory factor TNF-α in BV2 cells was detected using ELISA (n = 4). L The levels of the M1 phenotype-related inflammatory factor IL-6 in BV2 cells (n = 4). M The levels of the M2 phenotype-related anti-inflammatory factor IL-4 in BV2 cells (n = 4). N The levels of the M2 phenotype-related anti-inflammatory factor IL-10 in BV2 cells (n = 4). All experiments were performed in parallel. The results are presented as the mean ± SD. Statistical significance was determined using one-way ANOVA followed by an LSD post hoc test for multiple comparisons among the groups. *P < 0.05, **P < 0.01, ***P < 0.001, and NS, not significant.