Figure 4.
SARS-CoV-2 N protein inhibits the binding between TAK1 and TAB2, unlike SARS-CoV N protein. (A) HEK293T cells were transfected with Flag-TAK1, Flag-SARS-CoV-2 N, Flag-empty vector, HA-TAB2, and HA-empty vector. Lysates were collected for immunoprecipitation (IP) (anti-HA) and immunoblot (IB) analysis. The ratio of the gray value between IP TAK1-Flag and IP TAB2-HA was determined. (B) HEK293T cells were transfected with Myc-empty vector or Myc-SARS-CoV-2 N for 48 h, followed by TNF-α treatment for 4 h. Lysates were subjected to immunoprecipitation (anti-TAB2) and immunoblot analysis. The ratio of the gray value between IP TAK1 and IP TAB2 was determined. (C) Domain organization of TAB2 and its truncated mutants. CUE, Ubiquitin binding domain; TBD, TAK1 binding domain; NZF, NLP4-like zinc finger domain. (D–F) HEK293T cells were transfected Flag-empty vector, Flag-TAK1 (D), Flag-SARS-CoV-2 N (E), or Flag-SARS-CoV N (F) along with HA-TAB2 full-length (FL), HA-TAB2 (1–392aa), HA-TAB2 (1–496aa), HA-TAB2 (1–574aa), and HA-TAB2 (1–653aa). Lysates were subjected to immunoprecipitation (anti-Flag) and immunoblot analysis. (G) HEK293T cells were transfected with Flag-TAK1, Flag-empty vector, Myc-SARS-CoV N, Myc-empty vector, HA-TAB2, and HA-empty vector. Lysates were collected for immunoprecipitation (anti-HA) and immunoblot analysis. The ratio of the gray value between IP TAK1-Flag and IP TAB2-HA was determined. (H) HEK293T cells were transfected with Myc-empty vector or Myc-SARS-CoV N for 48 h, followed with TNF-α treatment for 4 h. Lysates were subjected to immunoprecipitation (anti-TAB2) and immunoblot analysis. The ratio of the gray value between IP TAK1 and IP TAB2 was determined. Data are done at least twice, and one representative is shown. Source data are available for this figure: SourceData F4.

SARS-CoV-2 N protein inhibits the binding between TAK1 and TAB2, unlike SARS-CoV N protein. (A) HEK293T cells were transfected with Flag-TAK1, Flag-SARS-CoV-2 N, Flag-empty vector, HA-TAB2, and HA-empty vector. Lysates were collected for immunoprecipitation (IP) (anti-HA) and immunoblot (IB) analysis. The ratio of the gray value between IP TAK1-Flag and IP TAB2-HA was determined. (B) HEK293T cells were transfected with Myc-empty vector or Myc-SARS-CoV-2 N for 48 h, followed by TNF-α treatment for 4 h. Lysates were subjected to immunoprecipitation (anti-TAB2) and immunoblot analysis. The ratio of the gray value between IP TAK1 and IP TAB2 was determined. (C) Domain organization of TAB2 and its truncated mutants. CUE, Ubiquitin binding domain; TBD, TAK1 binding domain; NZF, NLP4-like zinc finger domain. (D–F) HEK293T cells were transfected Flag-empty vector, Flag-TAK1 (D), Flag-SARS-CoV-2 N (E), or Flag-SARS-CoV N (F) along with HA-TAB2 full-length (FL), HA-TAB2 (1–392aa), HA-TAB2 (1–496aa), HA-TAB2 (1–574aa), and HA-TAB2 (1–653aa). Lysates were subjected to immunoprecipitation (anti-Flag) and immunoblot analysis. (G) HEK293T cells were transfected with Flag-TAK1, Flag-empty vector, Myc-SARS-CoV N, Myc-empty vector, HA-TAB2, and HA-empty vector. Lysates were collected for immunoprecipitation (anti-HA) and immunoblot analysis. The ratio of the gray value between IP TAK1-Flag and IP TAB2-HA was determined. (H) HEK293T cells were transfected with Myc-empty vector or Myc-SARS-CoV N for 48 h, followed with TNF-α treatment for 4 h. Lysates were subjected to immunoprecipitation (anti-TAB2) and immunoblot analysis. The ratio of the gray value between IP TAK1 and IP TAB2 was determined. Data are done at least twice, and one representative is shown. Source data are available for this figure: SourceData F4.

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