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Relation between <t>SMAD4</t> expression and sensitivity to Pro A of PC cells. (A) qPCR analysis of expression of indicated genes in PC cell lines. (B) Western blot analysis of SMAD4 protein in PC cell lines. (C) CCK-8 assay was used to determine the effects of SMAD4 on Pro A-induced cytotoxicity in PC cell lines. Each experiment was repeated three times. Error bars represent SEM. **, P<0.01; ***, P<0.001; ****, P<0.0001. PC, pancreatic cancer; Pro A, proscillaridin A; qPCR, quantitative polymerase chain reaction; CCK-8, cell counting kit-8; SEM, standard error of mean.
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Relation between SMAD4 expression and sensitivity to Pro A of PC cells. (A) qPCR analysis of expression of indicated genes in PC cell lines. (B) Western blot analysis of SMAD4 protein in PC cell lines. (C) CCK-8 assay was used to determine the effects of SMAD4 on Pro A-induced cytotoxicity in PC cell lines. Each experiment was repeated three times. Error bars represent SEM. **, P<0.01; ***, P<0.001; ****, P<0.0001. PC, pancreatic cancer; Pro A, proscillaridin A; qPCR, quantitative polymerase chain reaction; CCK-8, cell counting kit-8; SEM, standard error of mean.

Journal: Annals of Translational Medicine

Article Title: Proscillaridin A induces mitochondrial damage and autophagy in pancreatic cancer and reduces the stability of SMAD4 in Panc-1 cells

doi: 10.21037/atm-22-1085

Figure Lengend Snippet: Relation between SMAD4 expression and sensitivity to Pro A of PC cells. (A) qPCR analysis of expression of indicated genes in PC cell lines. (B) Western blot analysis of SMAD4 protein in PC cell lines. (C) CCK-8 assay was used to determine the effects of SMAD4 on Pro A-induced cytotoxicity in PC cell lines. Each experiment was repeated three times. Error bars represent SEM. **, P<0.01; ***, P<0.001; ****, P<0.0001. PC, pancreatic cancer; Pro A, proscillaridin A; qPCR, quantitative polymerase chain reaction; CCK-8, cell counting kit-8; SEM, standard error of mean.

Article Snippet: The coding DNA for the short hairpin RNA (shRNA) precursor targeting SMAD4 was inserted into pLKO.1 (Addgene), and co-transfected with packaging plasmids psPAX2 and pMD2.G (Addgene) into HEK293T cells for production of recombinant lentivirus.

Techniques: Expressing, Western Blot, CCK-8 Assay, Real-time Polymerase Chain Reaction, Cell Counting

Pro A reduces SMAD4 protein stability in Panc-1 cells. (A) Western blot analysis of SMAD protein level in Panc-1 treated with indicated concentrations of Pro A for 48 h. (B) Western blot analysis of SMAD protein levels in Panc-1 treated with DMSO or Pro A for indicated times. (C) Panc-1 cells were treated with DMSO or Pro A and CHX for the indicated time and SMAD4 protein levels were analyzed by western blot. (D) SMAD4 protein levels in Panc-1 cells treated with DMSO or Pro A and with or without MG132, was determined by western blot analysis. Normalized densitometry scanning results are shown under the blots. −, absence; +, presence. DMSO, dimethyl sulfoxide; Pro A, proscillaridin A; CHX, cycloheximide.

Journal: Annals of Translational Medicine

Article Title: Proscillaridin A induces mitochondrial damage and autophagy in pancreatic cancer and reduces the stability of SMAD4 in Panc-1 cells

doi: 10.21037/atm-22-1085

Figure Lengend Snippet: Pro A reduces SMAD4 protein stability in Panc-1 cells. (A) Western blot analysis of SMAD protein level in Panc-1 treated with indicated concentrations of Pro A for 48 h. (B) Western blot analysis of SMAD protein levels in Panc-1 treated with DMSO or Pro A for indicated times. (C) Panc-1 cells were treated with DMSO or Pro A and CHX for the indicated time and SMAD4 protein levels were analyzed by western blot. (D) SMAD4 protein levels in Panc-1 cells treated with DMSO or Pro A and with or without MG132, was determined by western blot analysis. Normalized densitometry scanning results are shown under the blots. −, absence; +, presence. DMSO, dimethyl sulfoxide; Pro A, proscillaridin A; CHX, cycloheximide.

Article Snippet: The coding DNA for the short hairpin RNA (shRNA) precursor targeting SMAD4 was inserted into pLKO.1 (Addgene), and co-transfected with packaging plasmids psPAX2 and pMD2.G (Addgene) into HEK293T cells for production of recombinant lentivirus.

Techniques: Western Blot