Screening differentially expressed genes in mouse hepatocarcinoma ascites cell line with high potential of lymphatic metastasis

Figure 1 Lymph node metastatic rates of Hca-F and Hca-P. A: Implanted tumor of Hca-F cells and Hca-P cells. H.E. ×400. (1) Implanted tumor of Hca-F cells. (2) Implanted tumor of Hca-P cells; B: Regional lymph node of implanted tumor of Hca-F cells and Hca-P cells. H.E. ×400. (1) Regional lymph node of implanted tumor of Hca-F cells. (2) Regional lymph node of implanted tumor of Hca-P cells.

Figure 2 High integrity and purity of total RNA. A: Total RNA of Hca-F and Hca-P cells. Lane 1: RNA from Hca-F cells; lane 2: RNA from Hca-P cells; M: DNA marker DL2000; B: mRNA of Hca-F and Hca-P cells. Lane 1: mRNA from Hca-F cells; lane 2: mRNA from Hca-P cells; M: DNA marker DL2000.

Figure 3 Effect of Rsa I digestion. Lanes 1, 3: cDNA of Hca-F and Hca-P cells; lane 2, 4cDNA of Hca-F and Hca-P cells after Rsa I digestion; M: DNA Marker DL2000.

Figure 4 Ligation efficiency analysis. Lane 1: Tester-1 as template, G3PDH3 “primer and PCR primer1; lane 2: Tester-1 as template, G3PDH3 “primer and G3PDH5 “primer; lane 3: Tester-2R as template, G3PDH3 “primer and PCR primer1; lane 4: Tester-1 as template, G3PDH3 “primer and G3PDH5 “primer.

Figure 5 Results of secondary PCR amplification. Lanes 1-3: Product of primary PCR amplification; lane 4: secondary PCR amplification product of PCR control cDNA; lane 5: secondary PCR amplification product of unsubtracted cDNA; lane 6: secondary PCR amplification product of subtracted cDNA, M: DNA Marker DL2000.

Figure 6 Analysis of subtration effect. PCR was performed on subtracted (lanes 1-4) or unsubtracted (lanes 5-8) secondary PCR products with G3PDH 5’ primer and 3’ primer. Lanes 1, 5: 20 cycles; Lanes 2, 6: 25 cycles; Lanes 3, 7: 30 cycles; Lanes 4, 8: 35 cycles; M: DNA marker DL2000.

Figure 7 Results of clone PCR amplication. There was an average insert size of 0.3-0.7 kb.