Skip to main content
. 2021 Nov 24;5(2):e202101010. doi: 10.26508/lsa.202101010

Figure S11. Transcriptional activity of Tcf/Lef is not required for phenotype switching of melanoma cells.

Figure S11.

(A) Immunoblotting analysis to assess the knockdown efficiencies against TCF4 and LEF1. M000921 proliferative melanoma cells were treated with siCtrl, siTCF4, siLEF1, or siLEF1/siTCF4 in the absence or presence of 2 d of TGFβ, Wnt-3a, or siLATS1/2 stimulation. Immunoblotting for TCF4 and LEF1 validated the efficient knockdown of these proteins. GAPDH was used as loading control. (B) Quantitative RT-PCR analysis to assess knockdown efficiencies (TCF4, LEF1, LATS1, and LATS2) as well as Wnt target gene expression (AXIN1, NOTUM, NKD1, and CTLA4) expression upon Wnt-3a treatment or siLATS1/2 or TGFβ-induced phenotype switching in M000921 cells. (C) Quantitative RT-PCR analysis of melanocyte (MITF) and mesenchymal (FN1 and ZEB1) marker gene expression upon upon Wnt-3a treatment or siLATS1/2 or TGFβ-induced phenotype switching in M000921 cells. (D) Quantitative RT-PCR analysis of YAP/TAZ (CYR61 and CTGF) and YAP/TAZ/SMAD (SERPINE1 and ANKRD1) target gene expression upon siLATS1/2 or TGFβ-induced phenotype switching in M000921 cells. Mean + SEM of n = 3 replicates are shown. *P < 0.05; **P < 0.01; ***P < 0.005; ratio-paired t test. For clarity, bars without asterisks indicate statistically nonsignificant. Red asterisks indicate the respective comparisons to siCtrl. Black asterisks indicate the comparisons as highlighted by bars.