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(D) CGNs transfected with shMCU or shvector, Flag-VDAC1 or empty vector as indicated were treated with 70 μmol/L H2O2 for 24 h.

Studies were carried out in small scale shake cultures with various combinations of different concentrations of inducer, induction at different optical densities (OD) and using either high or low copy expression vector, as indicated.

Briefly, HEK293 cells were transfected with HA-S6K1 wt, HA-S6K1 K100R, or vector as indicated.

These TRAF2 mutant constructs were transfected into HEK-293 cells along with MLK3 expression vector, as indicated in Fig. 4B.

The AhR-EYFP (enhanced yellow fluorescent protein) expression vector was produced by cloning the full-length murine AhR cDNA into the pEYFP vector as indicated [ 53].

This lack of activation was not because of a toxic effect of the pCMVhMdm2 (1 244) vector as indicated by the analysis of the control luciferase reporter activity, where hMdm2 (1 244) expression significantly increases this activity.

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A T-25 flask of HEK293T cells was transfected with 3 µg APOBEC3 plus 3 µg of wild-type or mutant Vif expression vectors as indicated.

20 hours after that, cells were transfected by a calcium phosphate protocol [39] using 2 to 6 µg of different NFAT5 expression vectors (as indicated) and an empty plasmid (pCMV-HA vector, Clontech) to normalize for the total amount of transfected DNA.

Moreover, it can be recapitulated in expression vectors, as indicated in Figure  6C.

Hek293T cells (7 × 10) were transfected with 5 μg of expression vectors as indicated.

(G ) 293T cells were transfected with expression vectors of Myc-Atg14, HA-ROC1, FLAG-ZBTB16, and Myc-Cul3 expression vectors as indicated and cultured for 24 hr.

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