Protein expression in mock- and SBCMV-infected brain pericytes. (A) Western blot showing expression of HCMV MIE and pp65 virion proteins in pericytes infected with SBCMV for 72 hours. Actin was the loading control. (B) Temporal expression of HCMV pp65 virion protein by real-time PCR in infected pericytes (SBCMV or Towne CMV) at 12, 24, 48, 72, and 96 hours postinfection. (C) Semi-quantitative RT-PCR for HCMV MIE, GAPDH, CXCL8/IL-8, RSAD2, CXCL11/I-TAC and CCL5/Rantes (72-hour exposure). GAPDH was the loading control. (D) Real time RT-PCR analysis for CCL5/Rantes, CXCL11/I-TAC, and CXCL8/IL-8 in pericytes 72 hours after SBCMV infection. This experiment was replicated three times, and amplifications were performed in triplicate and normalized to GAPDH. CCL5/Rantes, chemokine (C-C motif) ligand 5/Rantes; CXCL8/IL-8, blood–brain barrier; CXCL11/I-TAC, chemokine (C-X-C motif) ligand 11/I-TAC; GAPDH, gyceralaldehyde phosphate dehydrogenase; HCVM, human cytomegalovirus; MIE, major immediate early protein: pp65, human cytomegalovirus phosphorylated envelop protein expressed at late times during virus replication; RSAD2, radical S-adenosyl methionine domain-containing protein 2; RT-PCR, reverse transcription polymerase chain reaction; SBCMV, primary HCMV isolate from a patient.