elFigure2.GCASPCinhibitsGBC cell proliferation and tumorgrowth100Figure 2BE-,GnC-SD scramble1.5MockSGC-996GBC-SDSHRNA!aGC9SGCASPO3eavectorof04o80.5SGC-996nLEGBCSOCoMBeRsPevectSGC-996vectot..SGC-996GCASPC6PCNA业(29 kDa)ph1.000.980.241.001.143.473.0720.82oIB-Tublin(50 kDa)3010152025SGC-906SBOR.00DayspostinoculationDGCASPChRNA1shRNA2SGC-996SGC-996Z80-AvectorASP4040ki67joa203OO0"G0/G1AG0/G1sG2/MG2/Mki67FGBC-SDGBC-SD scrambleGBC-SD三scrambleGBC-SDShRNA(ol. paonscrambleInNGBC-SDShRNA-2GBC-SDshRNA-IGBC-SDShRNA-20-GBC-SD300101520255:shRNA-10.8-Dayspostinoculationa280.4XIPALOX0.2GBC-SDshRNA-20.5-90GBC-SDGBC-SDGBC-SDShRNA-ShRNA-290scrambleGBC-SDGRCIADGRCAD
Figure 2. GCASPC inhibits GBC cell proliferation and tumor growth
Figure 3.GCASPC binds to pyruvate carboxylase proteinure3BIS10SGC-996170PCSGC-996130NOZPCNOZ95D5'Antisense7253'GCASPCPBSKA1(1-472nt)55PBSK△2(1-233nt)43kDaEF1.0(oaoo)0.5-CU2nRNA0.0COXIVGCASPC0.5T0
Figure 3. GCASPC binds to pyruvate carboxylase protein
Figure 4. GCASPC decreases the protein level and activity of pyruvatecarboxylasebyinhibitingitsproteinstability.Figure4A20*t(uedBnl)KuaeeodBentPC10(130 kDa)1.871.001.651.000.38β-Tublin(50 kDa)GBC-SDSGC-996GBC-SDGBC-SDoceSARNA-1ShRNA-2scramblevectorBCDCHXMGI32DMSOcontrolPC(130kDa)1.000.310.41PCNAPCPC(29 kDa)0.451.920.161.002.251.001.901.002.531.000.340.30B-Tublirβ-TublinB-Tublin(50 kDa)ENA3.1-PCS1.5-scramblePeDCPCDNA3.1-PC+GCASPCGCASPC0-0PCDNA3.1-PC1.0PC1.000.422.091.090.5PCNA0.441.961.001.160.0-β-Tublin0n3
Figure 4. GCASPC decreases the protein level and activity of pyruvate carboxylase by inhibiting its protein stability
Figure 5. GCASPC is a target of miR-17-3p100Figure5BA1.5scrambleGCASPCshRNA-1ss1.25'-GAGTGCCAAAAACTGCAGA-3'GCASPC-wt-GCASPCshRNA-2scrambleCmiR-17-3p1.0m1.00.8miR-17-3p3-UUCACGGAAG--UGACGUCA-5"T0.6-萃8华de0.5GCASPC-mut5'-GATGTTAAAAAGTGTAGTA-3'0.00.01.5-GBC-SDOCUG-1GBC-SDOCUG-1**CeDS01.0-1.5scramble+miR-17-3p mimicsnw ayoog0.5-1.0PC30.50.01.000.52wtytmutASPC.3p-17-3PGCAS-aC0.0Cβ-Tublin0.51.01.52.02.53.03.50.0alpha-amanitin (hours)FE=-0.498,p=0.00110.1.5scramblessshRNA-GCASPC+miR-17-3pinhibitor8shRNA-GCASPC+miR-17-3pinhibitor1.06-40.5-2o.0.0161214234105048miR-17-3pexpression (μCt)
Figure 5. GCASPC is a target of miR-17-3p
上海交通大学医学院CasShanghai Jiao Tong University School of Medicinc细胞的传代与培养
细胞的传代与培养