Figure 5 | Scientific Reports

Figure 5

From: An epigenetic GPI anchor defect impairs TLR4 signaling in the B cell transdifferentiation model for primary human monocytes BLaER1

Figure 5

PIGH is epigenetically downregulated in GPI-negative BLaER1 cells. (a) Alignment of the PIGH locus RefSeq with the Sanger sequencing result for genomic DNA from the GPI-negative clones. Exon 2 is depicted in blue. The two nucleotides that are deleted in the PIGHP1 sequence are shown in red. (b) Two GPI-negative BLaER1 monoclonal cell lines (1, circle and 2, square) were treated with 0.4 µM (light red) or 2.0 µM (dark red) 5-aza-2′-deoxycytidine (5-aza) or left untreated (black) for 2, 4, and 7 days. The percentage of CD48- or CD55-positive cells among GFP-expressing cells was determined by FACS analysis. Shown is the mean + SD of data from n = 3 independent experiments. (c) After treatment of GPI-negative BLaER1 monoclonal cell lines with 0.4 µM 5-aza or leaving them untreated for 4 days, cells were transdifferentiated for 5 days. The percentage of CD14-positive cells among GFP-expressing cells was determined by FACS analysis. Shown is the mean + SD of data from n = 3 independent experiments. 5-aza-treated cells were compared to untreated control cells using unpaired t tests (**p ≤ 0.01, ***p ≤ 0.001).

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