Pestiviruses can be distinguished as two biotypes, cytopathogenic (cp) and noncytopathogenic (noncp), by the morphological changes that they induce during growth in cultured cells. In this study, the cp phenotype of several classical swine fever viruses (CSFV) was evaluated by the detections of the nonstructural proteins NS2-3 and NS3 using immunoprecipitation and Western blotting in different porcine cell lines. Most CSFVs that showed the exaltation of Newcastle disease virus (END) phenomenon (END⁺ viruses) did not induce cytopathic effect (CPE) in any cell line, and detections of NS2-3 and NS3 showed a strong signal for NS2-3 in the END⁺ virus-infected cells. However, clear CPE was observed in serum-free cultured cells (FS-L3 and CPK-NS) infected with viruses that induce intrinsic interference but did not show the END phenomenon (END^- viruses), and signal of NS3 was strongly detected than that of NS2-3 in these cells at 72 hr after infection. As the results of the analysis of FS-L3 cells infected with ALD (END⁺ virus) and ALD-END^- virus (END^- virus) at several incubations, the signal of NS3 detected was strengthened with CPE that become evident progressively. These results suggest that CPE is associated with the accumulation of NS3, which is promoted in serum-free cell lines infected with END^- viruses. Thus, indicating there is a close relationship between CPE and the quantity of NS3 produced in END^- CSFV infection.