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The hormone sensitivity of PbGR2b and PbGR1 differed in the transactivation assay, with PbGR2b having lower EC50 values and greater fold induction.
On closer examination, we found that the RP transcripts had the greater fold induction in G largely because of a greater repression during G starvation (See Discussion and Figure S1).
The genes encoding protein chaperones, LEA and dehydrin, were dramatically induced by water deficit by more than a 100-fold, with greater fold induction in the immature culm when compared to the mature culm.
In particular, our data suggest that stable receptor expression via lentiviral transduction or stable integration of expression vectors into safe harbors in host chromosomal DNA should result in both lower background and greater fold induction (Supporting Information).
The greater fold induction of luciferase expression from pGTLMIK compared to pGTRTL was due to the combination of lower basal and higher induced levels of expression with this vector.
Reporter activation was highest when the FRB domain was on the TC (fold induction = 10.2), but the background was lowest when the FRB domain was on the PC, and therefore, the fold induction was greater (fold induction = 13.4).
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Table 1 lists the 59 genes with the greatest fold induction as measured on the microarray.
The greatest fold induction (68-fold) was seen with a probe set identifying HMOX1, the gene that encodes heme oxygenase-1, the rate-limiting enzyme in heme catabolism.
The 12-copy genotype had the greatest fold induction of Hsp70 mRNA and the highest level of Hsp70 message after 30 and 60 minutes, but was not significantly different from the 6-copy genotype at these time points (Tables 2 and 3).
The top 50 highest nickel-induced genes (genes with greatest fold-induction by nickel treatment identified by RNA-Seq) with qualified H3K4me3 peaks within 5,000 bp flanking both sides of the TSSs are listed in Table 2. ChIP-Seq analysis was performed with an antibody specific for H3K4me3 on chromatin isolated from untreated and nickel-treated A549 cells after crosslinking with formaldehyde.
If we set a threshold for clinical steroid responsiveness to be a greater than 500 fold induction of mIL1R2 in vitro, then the sensitivity of this test is 100% with a confidence interval of 71.7 to 100%.
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