Fig. 8.

HeV infection but not HeV M protein expression induces DNA damage. a HeLa cells transfected to express GFP-HeV M protein without (DMSO) or with etoposide treatment were fixed 16 h p.t., immunostained for γH2AX phosphorylated at S139 (γH2AX-p) and imaged by CLSM. Images are representative of ≥15 fields of view. b Lysates of HEK-293T cells transfected to express GFP alone (empty vector (EV)) or GFP-HeV M protein and treated at 16 h p.t. without or with etoposide (3 h), were analyzed by IB using the indicated antibodies (data are representative of two independent experiments). c HeLa cells infected with HeV (MOI 5; which results in c. 100% cells infected) were fixed 24 h p.i. and immunostained for γH2AX-p prior to imaging by CLSM (images representative of ≥15 fields of view from three biological replicates). d Lysates of mock or HeV-infected HeLa cells were collected at 24 h p.i. and analyzed by IB using the indicated antibodies (data representative of three biological replicates). Scale bars correspond to 15 μm