Therefore, nanogold labeling of the different Mediator subunits conclusively validated the localizations of the Head and Middle modules.
This antibody could not be used to validate the localization of AdcA at the plasma membrane as it decorated non-specifically the plasma membrane of adcA knockout cells.
To further validate the localization of the Tail module, an oligohistidine tag was engineered onto the C terminus of Med14, which is a connector of the Middle and Tail module.
MpFISH validated also the localization of RP11-732m7 at 3p24-25, while the UCSC database [ 10] placed it at 3q26 (170.9–171.1 Mb).
A summary of the knockout strategy is given in Fig. 5A, B. We further validated the mitochondrial localization of CTMP in mouse tissue.
We have validated the tissue localization of selected tissue-specific genes identified with PAT-Seq by cloning their promoters and using them to drive expression of GFP in vivo.
We present below several lines of evidence that validate the localization of TC-Oleosin chimera in endoplasmic reticulum of the cells in producing intracellular sequestration of vitamin B12.
Consistent with the above findings, a comprehensive histological analysis revisiting potential astrocyte reactivity in post-mortem brains of HD patients showed the presence of astrogliosis in Grade 0 caudate nucleus and validated the co-localization of mHtt in astrocytes along with a grade-dependent reduction in GLT-1.
Thus, our method is gentle enough to allow the co-detection of genetically encoded fluorescent proteins simultaneous with immunostaining in transgenic embryos, facilitating co-localization studies to rapidly validate the proper localization of fluorescent fusion proteins.
After validating the mitochondrial localization of PaMT1-his in T4 and T5, copper levels in mitochondria of these transformants were determined (Fig. 4D).
