Fig. 5: LHPP inhibited stem cell-like characteristics in gastric cancer cells in vitro.

A Basic protein expression of LHPP in gastric cancer cell lines was detected by western blotting. B Stable LHPP-overexpressing AGS cells and LHPP-knockdown MKN45 cells were constructed. Western blotting confirmed the changes in LHPP expression. C Western blot analysis was used to detect the protein expression levels of CD44, NANOG, SOX2, and SOX9 in gastric cancer cells with LHPP knockdown or overexpression. D Immunofluorescence imaging shows the expression of LHPP and CD44 in AGS cells overexpressing LHPP. E Immunofluorescence imaging shows the expression of LHPP and CD44 in LHPP-knockdown MKN45 cells. F The impact of LHPP overexpression or knockdown on the growth of patient-derived gastric cancer organoids. Organoid diameters were quantitatively analyzed, and a Student’s t test was used to compare the differences between the two groups. G Immunofluorescence imaging shows the expression of LHPP and CD44 in patient-derived gastric cancer organoids with LHPP overexpression or knockdown. H Human phospho-kinase microarray analysis of conditioned medium from MKN45 cells with stable LHPP knockdown. The relative signal intensities of the indicated proteins are summarized below. I Spheroid formation was assessed in MKN45 cells transfected with shLHPP and treated with the GSK-3β inhibitor Laduviglusib (CHIR-99021; 10 µM). Student’s t test was used to compare the differences between the two groups. J Organoid size was assessed in patient-derived organoids transfected with LHPP knockdown and treated with the GSK-3β inhibitor Laduviglusib (CHIR-99021; 10 µM). Student’s t test was used to compare the differences between the two groups. K Western blot analysis was used to detect the protein expression levels of CD44, NANOG, SOX2, and SOX9 in patient-derived organoids transfected with LHPP knockdown or treated with the GSK-3β inhibitor. *p < 0.05; **p < 0.01; ***p < 0.001.