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. 2020 Mar 25;12(4):778. doi: 10.3390/cancers12040778

Figure 3.

Figure 3

Knockdown of YAP1 by siRNA affects the migration capacities in GB-d1 and G-415 gallbladder cancer cell lines. (A) Cytoplasmic and nuclear protein expression of YAP1 and phospho-YAP1 in GBC cell lines by Western blot and estimation of the YAP1 nuclear/cytoplasmic ratio. Normalized expression of cytoplasmic phospho-YAP1 and nuclear YAP1 was quantified by densitometry. (B) Immunocytochemistry of YAP1 in GBC cell lines (Magnification: 20×). GB-d1 and G-415 cancer cells were transfected with 25nM of siRNA against YAP1 or siRNA non-target control; (C) the knockdown of YAP1 was validated by assessing mRNA and protein levels in GB-d1 G-415 cell lines at 24 h. YAP1 was quantified by qRT-PCR using QARS and TFCP2 as internal controls, while α-tubulin was used as an internal control for loading protein. (D) Migration analysis of GB-d1 and G-415 gallbladder cancer cells treated for 24 h with siRNA non-target control and siRNA YAP1, n = 3. ** p < 0.01 and *** p < 0.001 by one-sample t-test. Magnification: 20×.