Fig. 2: Selective regulation of REL expression in TS cells.

a Selective reduction of Rel transcript levels in TS^KI4 cells relative to TS^WT cells. Transcripts of all NF-κB members were measured by qPCR. The data normalized to Rps11 are expressed as a fold-change relative to TS^WT cells and are the mean ± SEM of n = 5 biologically independent experiments. b Cytoplasm and nuclear protein levels of NFκB members were measured using Western blotting in TS^WT cells and TS^KI4 cells. Blots are representative of n = 3 biologically independent experiments. c, d Densitometry was used to quantify cytoplasmic (c) and nuclear (d) NF-κB protein expression (b) from three biologically independent experiments. Data are expressed as fold-change relative to TS^WT cells and are the mean ± SEM of n = 3 biologically independent experiments. e NFκB transcriptional activity is reduced in TS^KI4 cells relative to TS^WT cells. Luciferase activity was measured in cells expressing a NF-κB luciferase construct. Data shown are the fold change in luciferase activity relative to TS^WT cells and are the mean ± SEM of n = 3 biologically independent experiments. f, g NF-κB member transcript levels in undifferentiated TS cells (TS^WT), TS cells differentiated for 4 days (T^DIFF), and invasive trophoblasts (T^INV) were measured using qPCR. The data normalized to Gapdh are expressed as a fold-change relative to undifferentiated TS^WT cells and are mean ± range of n = 2 biologically independent experiments. **p-value < 0.01; ***p-value < 0.001, ****p-value < 0.0001; Student’s t test. (n.s. not significant). Western blots show cropped images. Uncropped images are available in Supplementary Materials.