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. 2018 Nov 8;47(1):283–298. doi: 10.1093/nar/gky1149

Figure 5.

Figure 5.

Transgenerational transmission of TNF-α-induced TERRA into zygotes via spermatozoa. (A) TNF-α increases TERRA level in spermatozoa in an ATF7-dependent manner. WT and Atf7+/– male F0 mice (n = 3) were treated with TNF-α or saline as described in Figure 1A. TERRA expression in sperm was determined by poly(A)-tailed qRT-PCR. The average value relative to U2 snRNA ± SD is shown (n = 3). **P < 0.01; ***P < 0.001; NS, not significant. Control data for poly(A)-tailed qRT-PCR are shown in Supplementary Figure S5. WT and Atf7+/– male F0 mice (n = 3) were administered with TNF-α or saline as described in Figure 1A. TERRA expression in mature spermatozoa was determined by poly(A)-tailed qRT-PCR. Average value relative to U2 snRNA ± SD is shown (n = 3). **P < 0.01; NS, not significant. (B) Total RNA from spermatozoa was used for Northern blotting with 32P-labeled C24. The distribution of signal amounts is drawn on the right. (C) Transgenerational transmission of TNF-α-induced TERRA into zygotes. Spermatozoa was prepared from the TNF-α or saline-injected mice, and used for in vitro fertilization with the oocytes from female mice. The level of TERRA in two-cell embryos was determined by RNA FISH. Box plots representing the fluorescence intensity (arbitrary units, a.u.) of individual TERRA foci from nuclei (n = 40) are shown. ***P < 0.001. Typical RNA FISH data are shown.