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. 2020 Jul 31;28(1):219–232. doi: 10.1038/s41418-020-0596-y

Fig. 4. LINC00941 inhibited the ubiquitination of SMAD4 by blocking the binding of SMAD4 to βTrCP.

Fig. 4

a The expression of SMAD4 and βTrCP in HCT-116 and LoVo cells transfected with LINC00941 or control siRNA was determined by Western blotting. β-Actin was used as a loading control. b Western blot analysis of ubiquitinated SMAD4 immunoprecipitated from HCT-116 and LoVo cells transfection with LINC00941 or control siRNA. Total lysates of LoVo cells with or without LINC00941 knockdown (c) or HCT-116 cells with or without LINC00941 overexpression (d) were subjected to IP with anti-SMAD4 or anti-βTrCP Ab, followed by Western blotting using the indicated antibodies (Abs). The cells were treated with MG132 to inhibit the proteasome. e Total lysates from HCT-116 cells overexpressing Flag-tagged full-length or truncated SMAD4 protein were subjected to IP with anti-Flag Ab, followed by Western blotting using the indicated antibodies (Abs). Ig heavy chain (H chain) was used as a loading control. Data are presented as the mean ± SD. *p < 0.05, **p < 0.01, ***p < 0.001.