Fig. 4.

LINC00629 inhibits KLF4 degradation. A-B MNNG/HOS and 143B cells with or without LINC00629 knockdown were treated with 20 μM of the proteasome inhibitor MG132 for 8 h. The expression levels of KLF4 were detected by Western blot. Numbers represent the relative intensities of western blot bands of KLF4 to GAPDH. C-D MNNG/HOS and 143B cells with or without LINC00629 knockdown were treated with 10 mg/ml cycloheximide (CHX) for the indicated times. The expression levels of KLF4 were detected by Western blot (C), and the quantification of KLF4 levels relative to GAPDH is shown (D). The results are shown as the mean ± s.d. n = 3 independent experiments. P = 0.0002. E-F) MNNG/HOS cells with or without LINC00629 overexpression were treated with 10 mg/ml cycloheximide (CHX) for the indicated times. The expression levels of KLF4 were detected by Western blot (E), and the quantification of KLF4 levels relative to GAPDH is shown (F). The results are shown as the mean ± s.d. n = 3 independent experiments. P = 0.003. G MNNG/HOS cells with or without LINC00629 overexpression were transfected with the indicated constructs. After 24 h, the cells were treated with 20 μM MG132 for 8 h before collection. The whole-cell lysates were subjected to immunoprecipitation with KLF4 antibody and Western blot with anti-Ub antibody to detect ubiquitylated KLF4. H MNNG/HOS cells with or without LINC00629 knockdown were transfected with the indicated constructs. After 24 h, the cells were treated with 20 μM MG132 for 8 h before collection. The whole-cell lysates were subjected to immunoprecipitation with KLF4 antibody and Western blot with anti-Ub antibody to detect ubiquitylated KLF4. I MNNG/HOS cells with or without LINC00629 knockdown were transfected with the indicated constructs and treated with 3 μM TM. The cells were treated with 20 μM MG132 for 8 h before collection. The whole-cell lysates were subjected to immunoprecipitation with KLF4 antibody and Western blot with anti-HA antibody to detect ubiquitylated KLF4