Journal: Antiviral research

Article Title: Aberrant regulation of the Akt signaling network by human cytomegalovirus allows for targeting of infected monocytes.

doi: 10.1016/j.antiviral.2018.07.015

Figure Lengend Snippet: Primary monocytes were mock infected, HCMV infected, GM-CSF treated, or M-CSF treated for 24 hours. Monocytes were also pretreated with MK (an AKT inhibitor) for one hour prior to a 24 h infection. Cells were then lysed and subjected to protein microarray analysis from Fullmoon Biosystems. Data are the mean from 3 independent blood donors.

Article Snippet: Mock infection was performed by adding an equivalent volume of RPMI 1640 medium to monocytes, while GM-CSF or M-CSF treatment was performed by adding an equivalent volume of RPMI 1640 medium with recombinant human GM-CSF or M-CSF at 100 ng/ml (R&D Systems, Minneapolis, MI).

Techniques: Infection, Microarray

Journal: Antiviral research

Article Title: Aberrant regulation of the Akt signaling network by human cytomegalovirus allows for targeting of infected monocytes.

doi: 10.1016/j.antiviral.2018.07.015

Figure Lengend Snippet: (A-B) Peripheral blood monocytes were mock or HCMV infected, or treated with GM-CSF or M-CSF for (A) 24 h or (B) 30 m. (C-D) Monocytes were pretreated with LY294002 (a PI3K inhibitor), MK, or rapamycin (a mTOR inhibitor) for 1 h. Following treatment with inhibitors, cells were mock or HCMV infected for (C) 24 h or (D) 30 min. (A-D) Levels of p-S6K (T389), and S6K were detected by immunoblotting from whole cell lysates. Membranes were then reprobed for β-actin as a loading control. Data are representative of 3–6 independent blood donors.

Article Snippet: Mock infection was performed by adding an equivalent volume of RPMI 1640 medium to monocytes, while GM-CSF or M-CSF treatment was performed by adding an equivalent volume of RPMI 1640 medium with recombinant human GM-CSF or M-CSF at 100 ng/ml (R&D Systems, Minneapolis, MI).

Techniques: Infection, Western Blot

Journal: Antiviral research

Article Title: Aberrant regulation of the Akt signaling network by human cytomegalovirus allows for targeting of infected monocytes.

doi: 10.1016/j.antiviral.2018.07.015

Figure Lengend Snippet: (A) Monocytes were mock or HCMV infected, or treated with GM-CSF or M-CSF for 24 h. (B) Monocytes were infected with HCMV strain TB40E or Towne/E for 24 h. (C-D) Monocytes were pretreated with LY, MK, or rapamycin for 1 h. Cells were then mock or HCMV infected for 24 h. (A-D) Levels of p-XIAP and XIAP were determined by immunoblotting. Membranes were then reprobed for β-actin as a loading control. Data are representative of 3–6 independent blood donors.

Article Snippet: Mock infection was performed by adding an equivalent volume of RPMI 1640 medium to monocytes, while GM-CSF or M-CSF treatment was performed by adding an equivalent volume of RPMI 1640 medium with recombinant human GM-CSF or M-CSF at 100 ng/ml (R&D Systems, Minneapolis, MI).

Techniques: Infection, Western Blot

Journal: Antiviral research

Article Title: Aberrant regulation of the Akt signaling network by human cytomegalovirus allows for targeting of infected monocytes.

doi: 10.1016/j.antiviral.2018.07.015

Figure Lengend Snippet: (A-B) Peripheral blood monocytes were mock or HCMV infected, or treated with GM-CSF or M-CSF for (A) 30 m or (B) 24 h. (C) Monocytes were pretreated with 4EGI-1 (an eIF4E inhibitor) for 1 h, then mock or HCMV infected for 24 h. Levels of Mcl-1, HSP27, and XIAP expression were determined by immunoblotting. (D-E) Monocytes were pretreated with LY294002, MK, or rapamycin for 1 h. Following pretreatment, cells were mock or HCMV infected for (D) 30 m or (E) 24 h. 4E-BP1 and p-4E-BP1 expression were determined by immunoblotting. Electrophoresis of 4E-BP1 separates into three different forms (Constantinou and Clemens, 2005). The γ band corresponds to the most highly phosphorylated form of 4E-BP1, whereas the β and α bands represent the intermediate and least phosphorylated form, respectively. (A-E) Membranes were reprobed for β-actin as a loading control. Data are representative of 4–6 independent blood donors.

Article Snippet: Mock infection was performed by adding an equivalent volume of RPMI 1640 medium to monocytes, while GM-CSF or M-CSF treatment was performed by adding an equivalent volume of RPMI 1640 medium with recombinant human GM-CSF or M-CSF at 100 ng/ml (R&D Systems, Minneapolis, MI).

Techniques: Infection, Expressing, Western Blot, Electrophoresis

Journal: Antiviral research

Article Title: Aberrant regulation of the Akt signaling network by human cytomegalovirus allows for targeting of infected monocytes.

doi: 10.1016/j.antiviral.2018.07.015

Figure Lengend Snippet: (A-B) Monocytes were mock or HCMV infected, or treated with GM-CSF or M-CSF for (A) 30 m or (B) 24 h. (C-D) Monocytes were pretreated with LY294002, MK, or rapamycin for 1 h, after which cells were either mock or HCMV infected for (C) 30 m or (D) 24 h. (A-D) HSF1 and p-HSF1 expression were determined by immunoblotting. (E-F) Prior to 24 h mock or HCMV infection, monocytes were pretreated with KRIBB11 (a HSF1 inhibitor) for 1 h. (E) mTOR and p-mTOR or (F) HSF1, p-HSF1, Mcl-1, HSP-27, and XIAP expression were assessed by immunoblot. (A-F) Membranes were then reprobed for β-actin as a loading control. Data are representative of 3–5 independent blood donors.

Article Snippet: Mock infection was performed by adding an equivalent volume of RPMI 1640 medium to monocytes, while GM-CSF or M-CSF treatment was performed by adding an equivalent volume of RPMI 1640 medium with recombinant human GM-CSF or M-CSF at 100 ng/ml (R&D Systems, Minneapolis, MI).

Techniques: Infection, Expressing, Western Blot