Figure 5.
DsRNA rather than other NSPs is critical for determining the DMV size. (A) IB analysis of the NSP12, NSP5, and NSP15 expression in HEK-293T cells stably expressing SARS-CoV-2 NSP3-16, where NSP5-11 and NSP13-16 were polyproteins. (B) Quantification of the size of DMVs is shown as mean ± SEM. n (number of analyzed DMVs) = 111, 339, and 401 for NSP3/4, NSP3-16, NSP3-16+dsRNA, respectively. Student’s two-tailed t test, ***: P < 0.001. ****: P < 0.0001. ns: not significant. (C) TEM analysis of the DMVs induced by SARS-CoV-2 at 6 or 24 hpi with or without RDV treatment. Each boxed area in a low-magnification TEM micrograph (left) is depicted in detail in a high-magnification one (right). Scale bars: 2 μm for low-magnification TEM micrographs, 200 nm for high-magnification TEM micrographs. (D) Quantification of the size of DMVs is shown as mean ± SEM (left). n (number of analyzed DMVs) = 129, 128, 255, and 47 for control 6 hpi, RDV 6 hpi, control 24 hpi, and RDV 24 hpi, respectively. Quantification of DMV density is shown as mean ± SEM (right). n (number of analyzed DMVs) = 10, 9, 10, and 10 for control 6 hpi, RDV 6 hpi, control 24 hpi, and RDV 24 hpi, respectively. Student’s two-tailed t test, **: P < 0.01. ****: P < 0.0001. ns: not significant. (E) TEM analysis of the DMVs induced by MHV-A59 at 4 or 8 hpi with or without RDV treatment. Each boxed area in a low-magnification TEM micrograph (left) is depicted in detail in a high-magnification one (right). Scale bars: 2 μm for low-magnification TEM micrographs, 200 nm for high-magnification TEM micrographs. (F) Quantification of the size of DMVs is shown as mean ± SEM (left). n (number of analyzed DMVs) = 84, 33, 88, and 56 for control 4 hpi, RDV 4 hpi, control 8 hpi, and RDV 8 hpi, respectively. Quantification of DMV density is shown as mean ± SEM (right). n (number of analyzed DMVs) = 22, 19, 43, and 27 for control 4 hpi, RDV 4 hpi, control 8 hpi, and RDV 8 hpi, respectively. Student’s two-tailed t test, **: P < 0.01. ***: P < 0.001. ****: P < 0.0001. ns: not significant. Source data are available for this figure: SourceData F5.

DsRNA rather than other NSPs is critical for determining the DMV size. (A) IB analysis of the NSP12, NSP5, and NSP15 expression in HEK-293T cells stably expressing SARS-CoV-2 NSP3-16, where NSP5-11 and NSP13-16 were polyproteins. (B) Quantification of the size of DMVs is shown as mean ± SEM. n (number of analyzed DMVs) = 111, 339, and 401 for NSP3/4, NSP3-16, NSP3-16+dsRNA, respectively. Student’s two-tailed t test, ***: P < 0.001. ****: P < 0.0001. ns: not significant. (C) TEM analysis of the DMVs induced by SARS-CoV-2 at 6 or 24 hpi with or without RDV treatment. Each boxed area in a low-magnification TEM micrograph (left) is depicted in detail in a high-magnification one (right). Scale bars: 2 μm for low-magnification TEM micrographs, 200 nm for high-magnification TEM micrographs. (D) Quantification of the size of DMVs is shown as mean ± SEM (left). n (number of analyzed DMVs) = 129, 128, 255, and 47 for control 6 hpi, RDV 6 hpi, control 24 hpi, and RDV 24 hpi, respectively. Quantification of DMV density is shown as mean ± SEM (right). n (number of analyzed DMVs) = 10, 9, 10, and 10 for control 6 hpi, RDV 6 hpi, control 24 hpi, and RDV 24 hpi, respectively. Student’s two-tailed t test, **: P < 0.01. ****: P < 0.0001. ns: not significant. (E) TEM analysis of the DMVs induced by MHV-A59 at 4 or 8 hpi with or without RDV treatment. Each boxed area in a low-magnification TEM micrograph (left) is depicted in detail in a high-magnification one (right). Scale bars: 2 μm for low-magnification TEM micrographs, 200 nm for high-magnification TEM micrographs. (F) Quantification of the size of DMVs is shown as mean ± SEM (left). n (number of analyzed DMVs) = 84, 33, 88, and 56 for control 4 hpi, RDV 4 hpi, control 8 hpi, and RDV 8 hpi, respectively. Quantification of DMV density is shown as mean ± SEM (right). n (number of analyzed DMVs) = 22, 19, 43, and 27 for control 4 hpi, RDV 4 hpi, control 8 hpi, and RDV 8 hpi, respectively. Student’s two-tailed t test, **: P < 0.01. ***: P < 0.001. ****: P < 0.0001. ns: not significant. Source data are available for this figure: SourceData F5.

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