In the 24-hr panel, 2 transcripts (Ndufab1 and Gfm1) showed no regulation with any of the tested substances.
The regulation of three of the transcripts selected from the 24-hr panel (Nubp1, Ndufab1, Gfm1) could not be confirmed by RT-PCR.
Substances that are estrogenic increased epithelial cell DNA synthesis at the 24-hr end point and regulated transcripts in the 2-hr panel relative to vehicle or a nonestrogenic chemical at the 2-hr end point.
At the 24-hr end point, regulation of transcripts in the 24-hr panel by long- versus short-acting estrogens was distinct, because short-acting estrogens exhibited impeded regulation of these transcripts.
PPT was not included in the short- versus long-acting 24-hr panel analysis, because it is an ERα selective agonist but has not been classified as short- or long-acting.
Most of the probes in each of the panels confirmed the expected patterns (see Supplemental Material, Figure S3), with the 2-hr probes indicating regulation relative to vehicle by E2, E3 or D3 (see Supplemental Material, Figure S3A) and the 24-hr panel confirming differential regulation by long- versus short-acting estrogens (see Supplemental Material, Figure S3B).
Of the 10 transcripts from the 2-hr panel we tested, 8 showed similar responses with E2, E3, or D3, but D3 did not significantly induce two of the transcripts (Cdkn1a and Stat5a) and induced Nup50 less robustly than did E2 or E3.
D2SC cells were treated with LPS (100 ng/ml) or vehicle for 6 hr (upper panel) or 18 hr (lower panel).
MDA-MB-231 cells were heat shocked for 20 min at 43°C (HS) or treated with 80 μM arsenite for 2 hr (right panel).
DOI: http://dx.doi.org/10.7554/eLife.00459.004 10.7554/eLiFigure59.005 Figure 1 figure supplement 2. Mup1-pHluorin was used to quantify surface abundance of Mup1 for a population of yeast cells at 26°C or following a shift to 38°C for 2 hr (bottom panel).
