List of Supplementary Figures and Tables Supplementary Material, Fig. S1: Confirmation that YY1 and NLRP7 directly interact in HEK293T cells. Supplementary Material, Fig. S2: NLRP7 expression levels in human cell lines Supplementary Material, Fig. S3: NLRP7 (panel A) and NLRP2 (panel B) expression levels in BeWo cells before and after NLRP7 knockdown. Supplementary Material, Fig. S4: (A) Bisulfite validation of regions with altered methylation in undifferentiated hESC cultures. (B) FBXO4 expression in BeWo cells. (C) ZFP42 expression in BeWo cells. Supplementary Material, Fig. S5: Methylation differences at imprinted DMRs. Supplementary Material, Fig. S6: Methylation levels at ZFP42 in HM tissue. Supplementary Material, Table S1: List of 864 probes with significantly altered methylation at F>50 (Excel file) Supplementary Material, Table S2: List of 234 gene regions with altered methylation at F>5 (Excel file) Supplementary Material, Table S3: Statistics of identified probes with significantly altered methylation

Supplementary Material, Fig. S1 : YY1 and NLRP7 directly interact in HEK293T cells. N-terminal Myc-tagged YY1 and N-terminal V5-tagged NLRP7 were co-expressed in HEK293T cells. Compared to immunoprecipitation (IP) with IgG (negative control), IP with anti-V5 followed by immunoblotting (IB) with anti-Myc (upper panel), as well as immunoprecipitation with anti-NLRP7, followed by immunoblotting with anti-YY1 (lower panel) confirmed that NLRP7 binds to YY1. IP V5IP IgGInput IB: Myc IP NLRP7IP IgGInput IB: YY1

Supplementary Material, Fig. S2: NLRP7 expression levels in various human cell lines. Quantitative RT-PCR was performed to compare levels of NLRP7 between various cell lines. Expression was standardized to GAPDH expression and ΔΔCt method was used to quantify relative expression. The BeWo choriocarcinoma cell line has the highest levels, followed by H9 human embryonic stem cells (H9 hESC) which have ~30% NLRP7 expression compared to BeWo cells, and HEK293T cells, which have undetectable levels.

AB Supplementary Material, Fig. S3: NLRP7 (panel a) and NLRP2 (panel b) expression levels in BeWo cells before and after NLRP7 knockdown. Cells were transiently transfected with Dharmacon ON-TARGET plus siRNA Pool (siNLRP7 KD ) or the Non-Targeting Pool (siNLRP7 NTP ). Quantitative RT-PCR was performed to amplify NLRP7 mRNA (a) or NLRP2 mRNA (b). Expression was standardized to GAPDH expression and the ΔΔCt method was used to quantify relative expression compared to untransfected cells (Mock). The siRNA pool efficiently knocked down NLRP7 expression by ~85%, but did not affect NLRP2 expression.

CpG Clones H9 hESC shNLRP7 SC shNLRP7 KD Supplementary Material, Fig. S4: A) Bisulfite validation of regions with altered methylation in undifferentiated hESC cultures. B) FBXO4 expression in BeWo cells. Expression level determined by quantitative real-time PCR of FBXO4 is not different in cells transfected with the siRNA pool compared to cells transfected with the scrambled siRNA and mock transfected cells C) ZFP42 expression in BeWo cells. Expression level determined by quantitative real-time PCR of ZFP42 is also not different in cells transfected with the siRNA pool compared to cells transfected with the scrambled siRNA and mock transfected cells A B C

Supplementary Material, Fig. S5: Methylation differences at imprinted DMRs. Results at A) PEG3, B) KCNQ1OT1, C) H19 and D) SNRPN are shown for undifferentiated and differentiated shNLRP7 KD, shNLRP7 SC, and untransduced hES cells showing no difference in DNA methylation at any of these loci. Each is shown as a graph with the methylation levels on the Y-axis and the individual probes in the region listed on the X-axis. A B C D

TP#1 TP#2 POC#1POC#2POC#3 Pt#1 Pt#2 Pt#3 Supplementary Material, Fig. S6: Methylation levels at ZFP42 in HM tissue. Bisulfite sequencing at ZFP42 in A) Term placenta B) Products of conception, C) Corresponding patients with HM. No difference in DNA methylation is noted at ZFP42 when comparing term placenta with HM tissue. A B C