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. 2021 Aug 11;12:4856. doi: 10.1038/s41467-021-25072-x

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© This is a U.S. Government work and not under copyright protection in the US; foreign copyright protection may apply 2021

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Fig. 5 — a Graph showing the relative fold change (log2 or linear) expression of DUX, ZSCAN4 and MERVL in untreated or auxin-treated LTR-RFP reporter ESC^CTCF-AID for the indicated days. Center line shows the mean. Data are shown by triplicate. p-values are shown from two-tailed unpaired t-tests. n.s. = non-significant. b Box and whisker plot showing the averaged relative fold change (log2) expression of seven 2C-associated genes (DUX, ZSCAN4, ZFP352, TCSTV3, SP110, TDPOZ1 and MERVLs) in untreated or auxin-treated at the indicated time points in LTR-RFP reporter control and ESC^CTCF-AID. ESC were infected with lentiviruses expressing control or shRNAs against ZSCAN4. Reactions were performed by triplicate in two independent experiments. Center line indicates the median, box extends from the 25^th to 75^th percentiles and whiskers show Min to Max values. p-values are shown from two-tailed paired t-tests. Group comparisons at Day 1 and Day 2 in ESC^CTCF-AID are non-significant. All comparisons in parental cells are non-significant. c HTI quantification of RFP⁺ cells in untreated or auxin-treated for four days LTR-RFP reporter control and ESC^CTCF-AID. ESC were infected with lentiviruses expressing control or shRNAs against ZSCAN4. Center lines indicate mean values. n = 500; p-values are shown from two-tailed unpaired t-test. All comparisons in parental cells are non-significant. d HTI quantification of RFP⁺ cells in untreated or auxin-treated for 24 h LTR-RFP reporter control and ESC^CTCF-AID. ESC harbor a DOX-inducible piggyBac (PB) construct expressing ZSCAN4C and were induced with 1 μg/mL DOX as indicated together with auxin for 24 h. Center lines indicate mean values. n = 1500; p-values are shown from two-tailed unpaired t-tests. e Schematic representation of the model inferred from the data presented here. In a–d, one representative experiment is shown but two independent experiments were performed with similar results. In c, d percentages of RFP⁺ cells above the threshold are indicated.