Fig. 8: Loss of IQGAP3 reduces β-catenin levels in NUGC3 and AGS cells.

A Immunoblot of NUGC3 wild-type and IQGAP3 CRISPR knock-out clones. Samples were subjected to SDS-PAGE (7.5%) followed by immunoblotting using the indicated antibodies. The relative immunoblot bands of β-catenin and β-actin (βcat/βact) were quantified by densitometry. B NUGC3 cells were lysed, and RNA collected were converted to cDNA and subjected to RT-PCR to quantify the amount of Wnt target genes CCND1 and MYC. Representative data were collected and are expressed as the mean ± SD from three independent experiments. Student’s t test was performed, with ∗p ≤ 0.05, ∗∗p ≤ 0.01, and ∗∗∗p ≤ 0.001. C XTT Cell Proliferation Assay of NUGC3 wild-type and IQGAP3 CRISPR knock-out clones. Representative data were collected and are expressed as the mean % growth ±SD from three independent experiments. Two-way ANOVA was performed, with ∗p ≤ 0.05, ∗∗p ≤ 0.01, and ∗∗∗p ≤ 0.001. D Clonogenic Assay of NUGC3 wild-type and IQGAP3 CRISPR knock-out clones. E Colony-Forming Efficiency (%) of NUGC3 wild-type and IQGAP3 CRISPR knock-out clones. F Immunoblot of AGS wild-type and IQGAP3 CRISPR knock-out clones. Image is best representative of three independent experiments. Samples were subjected to SDS-PAGE (7.5%) followed by immunoblotting using the indicated antibodies. The relative immunoblot bands of β-catenin and α-Tubulin (βcat/αTub) were quantified by densitometry. G AGS cells were lysed, and RNA collected were converted to cDNA and subjected to RT-PCR to quantify the amount of Wnt target genes CCND1 and MYC. Representative data were collected and are expressed as the mean ± SD from three independent experiments. Student’s t test was performed, with ∗p ≤ 0.05, ∗∗p ≤ 0.01, and ∗∗∗p ≤ 0.001. H XTT Cell Proliferation Assay of AGS wild-type and IQGAP3 CRISPR knock-out clones. Representative data were collected and are expressed as the mean % growth ±SD from three independent experiments. Two-way ANOVA was performed, with ∗p ≤ 0.05, ∗∗p ≤ 0.01, and ∗∗∗p ≤ 0.001. I Clonogenic Assay of AGS wild-type and IQGAP3 CRISPR knock-out clones. J Colony-Forming Efficiency (%) of AGS wild-type and IQGAP3 CRISPR knock-out clones.