Additional file 1 of Exosome circATP8A1 induces macrophage M2 polarization by regulating the miR-1-3p/STAT6 axis to promote gastric cancer progression

Additional file 1: Figure S1. The fundamental characteristics of circATP8A1. A The genomic composition and circular structure of circATP8A1. B The expression of circATP8A1 in cell lines was verified by qPCR. C After treating AGS cells with Actinomycin D, the relative residual amounts of circATP8A1 and linear ATP8A1 were detected by qRT-PCR at different time points. CircATP8A1 (Circular circATP8A1), mRNA ATP8A1 (linear mRNA ATP8A1). D & E The relative residual amounts of circATP8A1 and linear ATP8A1 in AGS and MKN-45 cells before and after RNase treatment were detected by qRT-PCR. F Agarose gel electrophoresis was used to detect the expression of circATP8A1 (Circular circATP8A1) and ATP8A1 mRNA (linear mRNA ATP8A1) in the cDNA and gDNA, with β-actin serving as the control. P value is determined by t-test for B, D, and E. * P < 0.05, ** P < 0.01, *** P < 0.001. NS, not significant. Figure S2. CircATP8A1 knockdown reduces proliferation and migration in MKN-45 cells. A The relative expression levels of circATP8A1 and linear ATP8A1 after circATP8A1 knockout in MKN-45 cells were detected by qRT-PCR. B The proliferative ability of MKN-45 cells after circATP8A1 knockdown was detected by CCK8 assay. C & D Representative images of clone formation and statistics of colony counts in MKN-45 cells with circATP8A1 knockdown. E & F Microscopic images and quantification of the migration and invasion of MKN-45 cells as described above. P value is determined by t-test for A, B, D, and F. * P < 0.05, ** P < 0.01, *** P < 0.001. NS, not significant. shNC, the knockdown-control group; sh1-circATP8A, circATP8A1 knockdown group1; sh2-circATP8A, circATP8A1 knockdown group2. Figure S3. CircATP8A1 overexpression increases proliferation and migration in SGC7901 cells. A The relative expression levels of circATP8A1 and linear ATP8A1 after circATP8A1 overexpression in SGC-7901 cells were detected by qRT-PCR. B The proliferative ability of SGC-7901 cells after circATP8A1 overexpression was detected by CCK8 assay. C & D Representative images of clone formation and statistics of colony counts in SGC-7901 cells with circATP8A1 overexpression. E & F Microscopic images and quantification of the migration and invasion of SGC-7901 cells as described above (E & F). P value is determined by t-test for A, B, D, and F. * P < 0.05, ** P < 0.01, *** P < 0.001. NS, not significant. OC, overexpression control group; OE, circATP8A1 ovexpression group. Figure S4. Identification of exosomes from gastric cancer cell lines. A The exosomes of gastric cancer cell lines (AGS, SGC-7901, and MKN-45) were detected by electron microscopy. B Nanoparticle tracking analysis of exosomes in gastric cancer cell lines (AGS, SGC-7901, and MKN-45). C Western Blot analysis of exosome markers CD81, TSG101, and the negative marker Calnexin in gastric cancer cell lines. Table S1. The primers for qRT-PCR. Table S2. Association of circATP8A1 expression with clinicopathological features in plasma of gastric cancer patients.