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. Author manuscript; available in PMC: 2019 May 7.
Published in final edited form as: Free Radic Biol Med. 2007 Dec 4;44(5):847–855. doi: 10.1016/j.freeradbiomed.2007.11.012

Figure 2. Inactive HO-1 protein also increases the HO-1 promoter activity.

Figure 2

(A) 3T3-HO-1/luc cells were transiently transfected with pCMV vector, HO-1 cDNA (HO-1) and an enzymatically inactive HO-1 mutant in which Histidine 25 was substituted with Alanine (HO-1 H-A). A representative pseudoimage of photon emission is shown in the upper panel. Western blot analysis showed the 40 KD transfected HO-1 fusion proteins and the 32 KD endogenous HO-1 protein. Actin was used for loading control. Quantitation of the photon emission is shown in the lower panel. (B) Wildtype and inactive HO-1 proteins were delivered into 3T3-HO-1/luc cells. A representative pseudoimage of photon emission is shown in the upper panel. A representative Western blot of HO-1 immunoreactive protein is shown in the middle panel. Quantitation of the photon emission is shown in the lower panel. UT: untransfected; HO-1: Cells were HO-1 protein delivered; mutant HO-1: cells were an inactive mutant HO-1 (His25Ala) protein delivered. Values are expressed as a fold of untransfected controls and are the mean ± standard error of four separate determinations. *p < 0.05 vs. UT. (C) A pseudoimage of the photon emission from 3T3-HO-1/luc cells after delivery of HO-1 protein of 12 hours. Negative controls are cells that did not receive any protein (UT) or cells where other proteins such as galatosidase (Gal), a FITC-labeled immunoglobulin (AB) or Pro-ject alone (Proj) were delivered. Quantitative luminescence intensity is shown below. Values are the mean ± standard error of four independent measurements. *P<0.05 vs. UT.