CVB3 induces an autophagic response at different times during CVB3 infection in HeLa cells. HeLa cells were infected with CVB3, cell lysates were collected at 1, 3, 5, 7, 9, 12 and 24 h.p.i., and mock-infected HeLa cells served as the sham control. LC3, p62, p-mTOR and viral capsid protein VP1 were examined by western blot analysis, and these samples were immunoblotted with an antibody to β-actin to illustrate equal protein loading. Compared with the sham group, the LC3-II/LC3-I ratio increased at 1 h.p.i. then peaked at 5 h.p.i. (p<0.05). p62 exhibited a decrease at 1, 3, 5, 7 and 9 h.p.i. (p<0.01). No changes were noted at 12 and 24 h.p.i. compared with the sham group. p-mTOR showed a decrease after 5 h.p.i. (p<0.05), which was more obvious at 12 and 24 h.p.i. VP1 increased as the time of infection increased. *P<0.05, compared with the sham group; **p<0.01, compared with the sham group. CVB3, coxsackievirus B3; h.p.i., hours post-inoculation; LC3, light chain 3; mTOR, mammalian target of rapamycin.