Original Articles
Copyright ©The Author(s) 1999.
World J Gastroenterol. Jun 15, 1999; 5(3): 199-208
Published online Jun 15, 1999. doi: 10.3748/wjg.v5.i3.199
Figure 1
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Figure 1 Plasmid map of phHSP90. Vector: pSP64(3 kb); Inserts: human HSP90β cDNA (2.5 kb); Insert site: Sma I of multiclonal enzyme restriction site of pSP64; Promoter: SP6; Drug resistance: Amp.
Figure 2
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Figure 2 Plasmid map of eukaryotic expression vector pcDNA 3. 1 (+). (5.4 kb).
Figure 3
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Figure 3 Construction of pcDNA-HSP90 of HSP90 antisense RNA eukaryotic expression vector.
Figure 4
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Figure 4 Nuclease restriction analysis of pcDNA-HSP90 of HSP90 anti-sense RNA eukaryotic expression vector. 1. phHSP90; 2. phHSP90/EcoRI; 3. HSP90/EcoRI-BamHI; 4. λDNA/EcoRI+Hind III marker (21227, 5148, 4268, 2027, 1375, 125bp); 5. pcDNA 3.1(+); 6. pcDNA 3.1 (+)/EcoRI+BamHI; 7. pcDNA-HSP90; 8. pcDNA-HSP90/EcoRI+BamH; 9. pcDNA-HSP90/EcoRI; 10 pcDNA-HSP90/BamHI.
Figure 5
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Figure 5 SGC7901, SGC7901/VCR, HCC7402 and Ec109 cells transfected with pcDNA-HSP90 of HSP90 antisense RNA constructs, selected by G418, positive clones were obtained and were named. a. AH-SGC7901; b. AH-SGC7901/VCR; c. AH-HCC7402; d. AH-Ec109.
Figure 6
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Figure 6 The expression of β-actin mRNA in AH-SGC7901 of pcDNA-HSP90 transfected cell line, SGC7901-pcDNA of pcDNA 3. 1(+) transfected cell line and its parental cell line SGC7901 by Dot blot. 1: SGC7901; 2: SGC7901-pcDNA; 3: AH-SGC701.
Figure 7
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Figure 7 The expression of HSP90 anti-sense RNA in pcDNA-HSP90 transfected, pcDNA 3. 1(+) transfected and the parental cell lines of SGC7901, SGC7901/VCR, HCC7402 and Ec109 by Dot blot. 1: SGC7901; 2: SGC7901/VCR; 3: HCC7402; 4: Ec109.
Figure 8
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Figure 8 Expression of hsp90 anti-sense RNA by RNase protection assay. M: PBR322 DNA/Hae III Marker; C1: Positive control, β -actin (20bp) and HSP90 probe (30bp); C2: negative control, probe plus RNase S1, no RNA. 1. AH-SGC7901/VCR; 2. SGC7901-pcDNA/VCR; 3. SGC7901/VCR; 4. AH-SGC7901; 5. SGC7901-pcDNA; 6. SGC7901; 7. AH-HCC7402; 8. HCC7402-pcDNA; 9. HCC7402; 10. AH-Ec109; 11. Ec109-pcDNA; 12. Ec109.
Figure 9
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Figure 9 The expression of HSP90 protein in pcDNA-HSP90 transfected cells. 1. SGC7901; 2. SGC7901-pcDNA; 3. AH-SGC7901; 4. SGC7901/VCR; 5. SGC7901/VCR-pcDNA; 6. AH-SGC7901/VCR; 7. HCC7402; 8. HCC7402pcDNA; 9. AH-HCC7402; 10. Ec109; 11. Ec109pcDNA; 12. AHEc109.
Figure 10
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Figure 10 Growth of gene transfected cell lines.