Skip to main content
. 2023 Nov 13;13:19750. doi: 10.1038/s41598-023-47111-x

Figure 5.

Figure 5

RNF2 control threshold and persistence of Wnt activation by regulating TCF7L1. (a) Western blot analysis of control HeLa cells and RNF2 knockdown stable HeLa cells. TCF7L1-HA (6 μg) plasmid was introduced into cells, which were then treated with LiCl (25 mM, 50 mM) conditioned media for 16 h. Cells were lysed with RIPA buffer. (b) Western blot analysis of RNF2 knockdown stable HeLa cells. Cells were treated with LiCl (25 mM, 50 mM, and 100 mM) conditioned media for 16 h. Cells were lysed with RIPA buffer. ABC refers to active β-catenin. (c) Western blot analysis of control HeLa cells and RNF2 knockdown stable HeLa cells. TCF7L1-HA (6 μg) plasmid was introduced into cells, which were then treated with LiCl (25 mM, 50 mM, and 100 mM) conditioned media for 16 h. Cells were lysed with RIPA buffer. (d) RT-PCR analysis of total RNAs extracted from control HeLa cells and RNF2 knockdown stable HeLa cells after treatment with LiCl (50 mM) conditioned media for 8, 16, and 24 h. Samples were subjected to cDNA synthesis and RT-PCR analysis. (e) Western blot analysis of control HeLa cells and RNF2 knockdown stable HeLa cells treated with LiCl (25 mM and 50 mM) and CHIR99021 (5 μM and 10 μM) conditioned media for 16 h. TCF7L1-HA (6 μg) plasmid was introduced into control HeLa cells. (f) Western blot analysis of control 4T1 cells treated with LiCl (25 mM and 50 mM) and CHIR99021 (5 μM and 10 μM) conditioned media for 16 h. (g) Western blot analysis of control HEK239T cells and 4T1 cells treated with LiCl (25 mM, 50 mM, and 100 mM) and conditioned media for 16 h. (h) Western blot analysis of control HeLa cells and RNF2 knockdown stable HeLa cells treated with 50 mM LiCl conditioned media for varying durations (30 min, 1 h, 2 h, 4 h, 8 h, 16 h, and 24 h).