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. 2010 May 26;84(15):7898–7903. doi: 10.1128/JVI.00885-10

FIG. 4.

FIG. 4.

Increased abundance of cytosolic Grp75 in the heavy MAM fraction of HCMV-infected HFFs at 72 hpi. HFFs (P-13) were not infected or infected with HCMV (strain AD169, multiplicity of infection of 3) and harvested at 72 hpi. (A) The banding patterns of the MAM and mitochondria of uninfected and HCMV-infected HFFs in Percoll gradients. Uninfected and HCMV-infected HFFs (5 T175 flasks each) were fractionated in Percoll gradients as described previously (7, 8). The banding patterns of the MAM and mitochondria in the gradients are indicated. (B) Western analyses of uninfected and HCMV-infected HFFs. Ten-microgram amounts of total lysate and microsomal, cytosolic, heavy MAM, light MAM, and mitochondrial fractions were resolved by SDS-PAGE, blotted, and reacted with anti-UL37x1 antiserum (top panels) and DPM1 (1:200), COX2 (1:200), and Grp75 (1:2,500) antibodies. The positions of pUL37x1 (arrow) and a smaller UL37 species (asterisk) are indicated beside the blot. (C) Proteinase K sensitivity of cytosolic Grp75 in the heavy MAM fraction of HCMV-infected HFFs. Twenty-microgram amounts of the indicated fractions were not treated (−) or treated with proteinase K (+) as described for Fig. 3C and analyzed for the presence of Grp75 and pUL37x1.