Figure 7.
Figure 7. K63-linked ubiquitination is required for the formation of NEMO-containing structures in response to IL-1, but not TNF. U2OS-shUb-UbWT and U2OS-shUb-UbK63R cells cultured in the absence (−Dox.) or presence (+Dox.) of Dox (1 µg/ml) for 3 d were stimulated with IL-1 (A) or TNF (C), as indicated. Immunofluorescence assays were performed to detect NEMO. In C, cells were permeabilized with saponin before fixation and immunofluorescence assays. Nuclei were stained with DAPI (blue). Bars, 10 µm. (B and D) Numbers of NEMO spots per cell in the experiments presented in A and C, respectively, presented as mean values ± SD (error bars; n > 50 cells analyzed). Unpaired t tests were used to assess the significance of differences. ***, P < 0.001.

K63-linked ubiquitination is required for the formation of NEMO-containing structures in response to IL-1, but not TNF. U2OS-shUb-UbWT and U2OS-shUb-UbK63R cells cultured in the absence (−Dox.) or presence (+Dox.) of Dox (1 µg/ml) for 3 d were stimulated with IL-1 (A) or TNF (C), as indicated. Immunofluorescence assays were performed to detect NEMO. In C, cells were permeabilized with saponin before fixation and immunofluorescence assays. Nuclei were stained with DAPI (blue). Bars, 10 µm. (B and D) Numbers of NEMO spots per cell in the experiments presented in A and C, respectively, presented as mean values ± SD (error bars; n > 50 cells analyzed). Unpaired t tests were used to assess the significance of differences. ***, P < 0.001.

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