Porcine circovirus 2 (PCV2) infection activated calcium/calmodulin-dependent protein kinase kinase-beta (CaMKKβ) and its substrates calcium/calmodulin-dependent protein kinase I (CaMKI) and 5′ adenosine monophosphate-activated protein kinase (AMPK), as well as increased lipidation of microtubule-associated microtubule-associated protein 1 light chain 3 (LC3-II). PK-15 cells were infected with PCV2 (multiplicity of infection (MOI) ≈ 1) or mock-infected for the indicated time points. Whole cell lysates were subjected to Western blotting for CaMKKβ, phosphorylated AMPK (p-AMPK), total AMPK (t-AMPK), phosphorylated CaMKI (p-CaMKI), total CaMKI (t-CaMKI), viral capsid protein (Cap), and LC3-II. (A) Representative images of Western blotting for target proteins of cells at 12, 24, 36 and 48 hours post-infection (hpi); (B) ratios of CaMKKβ and p-AMPK to β-actin; (C) ratios of p-CaMKI and LC3-II to β-actin. Ratios of targeted proteins to β-actin were normalized to mock infection set at 1.0. Data are reported as the mean ± SEM of three independent experiments (ns, p > 0.05; * p < 0.05; and ** p < 0.01).