We would like to thank Dr. Stephane Gourguechon and Dr. Zacheus Cande, University of California Berkeley, for providing the plasmid construct pc-3HABSR.
The resulting construct, pC-TbAUK1-PTP-NEO, was linearized at the unique Nru I restriction site, and transfected into the procyclic cell line 427.
The resulting construct pc-GlLa3HABSR was linearized at the Eco47III site located in the middle of the coding region of GlLa (Figure S1) and electroporated into Giardia WB strain (WB clone C6, ATCC 50803) trophozoites for homologous recombination between the 3' half of the 3X-HA tagged GlLa gene and one of the endogenous GlLa genes [25], [26].
To generate cassettes for inactivation of the KU70 and KU80 genes, left flanks (LF) and right flanks (RF) of each gene were cloned by yeast recombinational cloning on either side of the resistance marker in the vectors pC-G418-YR and pC-SUR-YR to construct pC-G418-KU70-KO, pC-G418-KU80-KO and pC-SUR-KU80-KO (supplementarytary materials and methods).
To investigate whether the DFO-induced increase in p-AKT was dependent on PTEN levels, we incubated PC-3 cells stably transfected with the vector alone (PC-3-Control) or a PTEN construct (PC-3-PTEN) with DFO or Dp44mT (Supplementary Figure 5A and B).
The loadings matrix, P, describes the contributions of each variable to the constructed PCs.
However, detailed comparison of the constructed pseudo Pc curves with the experimented results illustrates that the constructed Pc curves are overestimated.
Digital realizations sampled from the constructed PC representations based on both schemes capture the observed statistical characteristics of the experimental data with sufficient accuracy.
The parts for constructed Pc curves with mercury injection saturations (SHgs) higher than 35% are all under the laboratory experimented results.
In particular, the performance of PSSMs converted from HMMs (PO) versus those iteratively constructed (PC and PS) shows that a more carefully constructed profile may yield better performance, with the difference being more pronounced in SUPERFAMILY than in Pfam.
An alternative approach would be to construct one PC to represent each functional group (for example, one temperature PC, one precipitation PC, etc).
