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EVs from ΔICP0-infected cells are markedly different than EVs released from HSV-1(F)-infected cells. (A and B) Total EVs were isolated from the supernatant of uninfected, HSV-1(F)-infected, or ΔICP0-infected hTERT-HEL cells at 48 h post-infection (0.5 PFU/cell). Supernatant was filtered and concentrated as in Fig. 1, EVs were then pelleted at 100,000 × g for 1 h and total EVs were loaded for immunoblot analysis using antibodies against CD63, <t>TSG101,</t> CIN85, LC3, Rab11, ARF6, and Annexin A1. Equal amounts of proteins from total cell lysates were analyzed by western blot for the same targets. ICP0 was used a control for infection and β-actin as a loading control.
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EVs from ΔICP0-infected cells are markedly different than EVs released from HSV-1(F)-infected cells. (A and B) Total EVs were isolated from the supernatant of uninfected, HSV-1(F)-infected, or ΔICP0-infected hTERT-HEL cells at 48 h post-infection (0.5 PFU/cell). Supernatant was filtered and concentrated as in Fig. 1, EVs were then pelleted at 100,000 × g for 1 h and total EVs were loaded for immunoblot analysis using antibodies against CD63, <t>TSG101,</t> CIN85, LC3, Rab11, ARF6, and Annexin A1. Equal amounts of proteins from total cell lysates were analyzed by western blot for the same targets. ICP0 was used a control for infection and β-actin as a loading control.
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EVs from ΔICP0-infected cells are markedly different than EVs released from HSV-1(F)-infected cells. (A and B) Total EVs were isolated from the supernatant of uninfected, HSV-1(F)-infected, or ΔICP0-infected hTERT-HEL cells at 48 h post-infection (0.5 PFU/cell). Supernatant was filtered and concentrated as in Fig. 1, EVs were then pelleted at 100,000 × g for 1 h and total EVs were loaded for immunoblot analysis using antibodies against CD63, <t>TSG101,</t> CIN85, LC3, Rab11, ARF6, and Annexin A1. Equal amounts of proteins from total cell lysates were analyzed by western blot for the same targets. ICP0 was used a control for infection and β-actin as a loading control.
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Average 94 stars, based on 1 article reviews
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EVs from ΔICP0-infected cells are markedly different than EVs released from HSV-1(F)-infected cells. (A and B) Total EVs were isolated from the supernatant of uninfected, HSV-1(F)-infected, or ΔICP0-infected hTERT-HEL cells at 48 h post-infection (0.5 PFU/cell). Supernatant was filtered and concentrated as in Fig. 1, EVs were then pelleted at 100,000 × g for 1 h and total EVs were loaded for immunoblot analysis using antibodies against CD63, TSG101, CIN85, LC3, Rab11, ARF6, and Annexin A1. Equal amounts of proteins from total cell lysates were analyzed by western blot for the same targets. ICP0 was used a control for infection and β-actin as a loading control.

Journal: Journal of Virology

Article Title: Effects on exocytosis by two HSV-1 mutants unable to block autophagy

doi: 10.1128/jvi.00757-23

Figure Lengend Snippet: EVs from ΔICP0-infected cells are markedly different than EVs released from HSV-1(F)-infected cells. (A and B) Total EVs were isolated from the supernatant of uninfected, HSV-1(F)-infected, or ΔICP0-infected hTERT-HEL cells at 48 h post-infection (0.5 PFU/cell). Supernatant was filtered and concentrated as in Fig. 1, EVs were then pelleted at 100,000 × g for 1 h and total EVs were loaded for immunoblot analysis using antibodies against CD63, TSG101, CIN85, LC3, Rab11, ARF6, and Annexin A1. Equal amounts of proteins from total cell lysates were analyzed by western blot for the same targets. ICP0 was used a control for infection and β-actin as a loading control.

Article Snippet: The mouse monoclonal antibodies to ICP0, CD63, Rab11, CIN85, NPC2, LAMP1, Rho A, ATG5, Annexin V, TSG101 (Santa Cruz), Flotillin-2 (BD Biosciences), β-actin (Sigma), p62/SQSTM1, Alix (Cell Signaling Technologies), and ARF6 (Sigma) were used in a 1:1,000 dilution.

Techniques: Infection, Isolation, Western Blot