We expressed Rv3802 in Escherichia coli and purified the enzymatically active form.
We expressed ANAC042-TEV-IFP-6xHis ANAC042-TEV-IFP-6xHis ANAC042-TEV-IFP-6xHis ANAC042-TEV-IFP-6xHis
When we expressed mRFP-Cdc42V12, inducedced the translocation of full-length APC to the leading edge in HCT116 cells as seen by a line intensity analysis (Fig. 5 ii) panels a, b and e) but mRFP-Cdc42N17 did not (Fig. 5 ii) panel d).
Specifically, we expressed BA0485 and BA2446 proteins containing a carboxyl-terminal hex-histidine tag from pET30-based vectors in E. coli BL21 DE3) Codon Plus-RIL induced with 1 mM IPTG.
These reports can be considered encouraging, but still leave open the question of whether parsing data into more specific subsets is worthwhile, despite the hopes we expressed last year on this count.
Following knockdown of endogenous Orai1 protein, we expressed V102C-Orai1 channels.
Using a third independent transgenic line, we expressed hMeCP2Δ166, which lacks the N-terminus including the MBD (Fig. 4C) (Cukier et al., 2008), via the alrm-Gal4 driver.
Therefore, we expressed Cdc8pE104A and constructed cdc8 E104A strains (SH41, SH104, SH105) because of the possibility to identify a fungal-specific function for this conserved residue.
As an independent verification of the data, we expressed mCherry-RTN1523, alore or in the presence of the RyR2 receptor in HEK293 cells, and examined the subcellular distribution of the RTN1523 construct.
In the present study, we expressed Rv2131c heterologously and found that the protein dephosphorylates PAP in a magnesium-dependent manner, with optimal activity observed between pH 8.5 and pH 9.5 using 0.5 mM MgCl2.
To test whether the activation of NFAT following store depletion in cells expressing V102C-Orai1 channels was indeed due to channel localization in ER-PM junctions, we expressed V102C-Orai1 channels in which L273 had been mutated L273S-Orai11) together with STIM1 and NFAT1-cherry in HEK cells in which Orai1 had been knocked down.
