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The protein showed cellular internalization and superior protection against paraquat-induced cell death as compared to either complex bi-functional protein without TAT or to native enzymes fused with TAT.
In the CA20948 tumour-bearing animals without TAT, a median survival of 29 days was found for animals bearing large tumours, whereas for animals bearing small tumours, this was 35 days.
Cells internalized TAT-Unc119 efficiently compared to Unc119 without TAT (Figure S5C).
In the case of luciferase assays without Tat treatment, co-transfection with a Renilla luciferase reporter driven by the constitutively active herpes simplex virus type 1 thymidine kinase (HSV-1 TK) promoter was used as an internal control for transfection efficiency.
Without Tat, AFF4 can mediate the ELL2-P-TEFb interaction, albeit inefficiently.
Although there may be some bias due to exclusion of children without Tat information, sensitivity analysis suggest that this did not influence our conclusions.
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TAT-Ndi1 increased the ratio 3-fold over I/R conditions without TAT-Ndi1 (Figure 5D).
To detect mitochondrial damage in adult cardiomyocytes subjected to sI/R with or without TAT-Ndi1, cytochrome c release was monitored by immunofluorescence.
Rat hearts were perfused 20 min with or without TAT-Ndi1 prior to 30 min global no-flow ischemia and 2 hr reperfusion.
All recombinant proteins were synthesized as mature proteins without the TAT signal peptide.
The extracellular add of 3 µM recombinant DJ-1 (a purified form of DJ-1 without the TAT sequence) had no such effect, confirming that only TAT-DJ-1 entered the plasma membrane and protected the cells [Figure 2G].
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