Fig 5.
Concurrent overexpression of nuclease-deficient ENDOG and EXOG inhibits mtDNA depletion by a mutant UL12.5-SPA protein. (A) Expression of myc-tagged constructs in transfected HeLa cells was visualized at 24 hpt. Lysates were assessed for protein expression by immunoblotting with the rabbit anti-c-myc antibody and chemiluminescence detection. (B) HeLa cells were cotransfected for 48 h with empty vector (pcDNA3.1) or plasmids expressing myc-tagged ENDOG, nuclease-deficient ENDOG (ENDOG-H141A), EXOG, nuclease-deficient EXOG (EXOG-H140A), or inactive sirtuin 3 (Sirt3-H248Y) along with an empty vector (pMZS3F) or a plasmid expressing UL12M185-G336A/S338A-SPA. MtDNA depletion was measured as described in the Fig. 3 legend. Data are from four separate experiments, except those data that include the Sirt3 mutant, which are from three separate experiments. All data were averaged, and standard errors are indicated. Data denoted with an asterisk (*) indicate statistical significance (P < 0.05) compared to the control data (UL12M185-G336A/S338A-SPA/pcDNA) as determined using a two-tailed t test with equal variances.