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primary antibodies against cyclin-dependent protein kinase 2  (Servicebio Inc)
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Servicebio Inc primary antibodies against cyclin-dependent protein kinase 2
RT-PCR and Western blotting validated differentially expressed miRNAs involved in the Wnt signaling pathway caused by GHaK treatment and expressions of their target genes. ( A ). RT-PCR analysis confirmed up-regulations of miR-4516, miR-4284, and miR-204-5p in PC-9 ( t = −18.74, −16.26, and −18.07, respectively, p < 0.01) and A549 cells ( t = −12.00, −17.64, and −9.70, respectively, p < 0.01) with additional up-regulations of miR-12136, miR-4463, and miR-1296-3p in PC-9 cells ( t = −8.70, p < 0.01; t = −4.32, −4.24, p < 0.05) caused by GHaK treatment. ( B ). RT-PCR analysis confirmed down-regulations of the target genes Wnt 8B, DVL3, and FOSL1 in both PC-9 ( t = 6.83, 7.98, and 5.30, respectively, p < 0.01) and A549 cells ( t = 5.68, 6.49, p < 0.01; t = 4.49, p < 0.05) with an additional down-regulation of FZD2 in PC-9 cells ( t = 5.00, p < 0.01) involved in the Wnt signaling pathway caused by GHaK treatment. ( C ). Western blotting analysis confirmed down-regulation expressions of p-GSK-3β, cyclinA1, and <t>CDK2</t> in A549 cells ( t = 19.46, 46.00, and 44.50, respectively, p < 0.01) and p-GSK-3β, cyclinB1, and CDK1 in PC-9 cells ( t = 25.36, p < 0.01; t = 4.52, p < 0.05; t = 72.00, p < 0.01) involved in the Wnt signaling pathway caused by GHaK treatment. (* p < 0.05; ** p < 0.01).
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RT-PCR and Western blotting validated differentially expressed miRNAs involved in the Wnt signaling pathway caused by GHaK treatment and expressions of their target genes. ( A ). RT-PCR analysis confirmed up-regulations of miR-4516, miR-4284, and miR-204-5p in PC-9 ( t = −18.74, −16.26, and −18.07, respectively, p < 0.01) and A549 cells ( t = −12.00, −17.64, and −9.70, respectively, p < 0.01) with additional up-regulations of miR-12136, miR-4463, and miR-1296-3p in PC-9 cells ( t = −8.70, p < 0.01; t = −4.32, −4.24, p < 0.05) caused by GHaK treatment. ( B ). RT-PCR analysis confirmed down-regulations of the target genes Wnt 8B, DVL3, and FOSL1 in both PC-9 ( t = 6.83, 7.98, and 5.30, respectively, p < 0.01) and A549 cells ( t = 5.68, 6.49, p < 0.01; t = 4.49, p < 0.05) with an additional down-regulation of FZD2 in PC-9 cells ( t = 5.00, p < 0.01) involved in the Wnt signaling pathway caused by GHaK treatment. ( C ). Western blotting analysis confirmed down-regulation expressions of p-GSK-3β, cyclinA1, and CDK2 in A549 cells ( t = 19.46, 46.00, and 44.50, respectively, p < 0.01) and p-GSK-3β, cyclinB1, and CDK1 in PC-9 cells ( t = 25.36, p < 0.01; t = 4.52, p < 0.05; t = 72.00, p < 0.01) involved in the Wnt signaling pathway caused by GHaK treatment. (* p < 0.05; ** p < 0.01).

Journal: Molecules

Article Title: Temporin-GHaK Exhibits Antineoplastic Activity against Human Lung Adenocarcinoma by Inhibiting the Wnt Signaling Pathway through miRNA-4516

doi: 10.3390/molecules29122797

Figure Lengend Snippet: RT-PCR and Western blotting validated differentially expressed miRNAs involved in the Wnt signaling pathway caused by GHaK treatment and expressions of their target genes. ( A ). RT-PCR analysis confirmed up-regulations of miR-4516, miR-4284, and miR-204-5p in PC-9 ( t = −18.74, −16.26, and −18.07, respectively, p < 0.01) and A549 cells ( t = −12.00, −17.64, and −9.70, respectively, p < 0.01) with additional up-regulations of miR-12136, miR-4463, and miR-1296-3p in PC-9 cells ( t = −8.70, p < 0.01; t = −4.32, −4.24, p < 0.05) caused by GHaK treatment. ( B ). RT-PCR analysis confirmed down-regulations of the target genes Wnt 8B, DVL3, and FOSL1 in both PC-9 ( t = 6.83, 7.98, and 5.30, respectively, p < 0.01) and A549 cells ( t = 5.68, 6.49, p < 0.01; t = 4.49, p < 0.05) with an additional down-regulation of FZD2 in PC-9 cells ( t = 5.00, p < 0.01) involved in the Wnt signaling pathway caused by GHaK treatment. ( C ). Western blotting analysis confirmed down-regulation expressions of p-GSK-3β, cyclinA1, and CDK2 in A549 cells ( t = 19.46, 46.00, and 44.50, respectively, p < 0.01) and p-GSK-3β, cyclinB1, and CDK1 in PC-9 cells ( t = 25.36, p < 0.01; t = 4.52, p < 0.05; t = 72.00, p < 0.01) involved in the Wnt signaling pathway caused by GHaK treatment. (* p < 0.05; ** p < 0.01).

Article Snippet: Polyvinylidene difluoride membranes (Servicebio, Cat No: G6015-0.45) were blocked with 5% non-fat milk in TBST buffer; incubated with primary antibodies against GAPDH (1:2000, Servicebio, Cat No: GB15004), ACTIN (1:2000, Servicebio, Cat No: GB15003), p-GSK-3β (1:1000, Servicebio, Cat No: GB114582), cyclin A1 (1:500, ABCLONAL, Cat No: A14529), cyclin-dependent protein kinase 1 (1:1000, CDK1, Servicebio, Cat No: GB11398), cyclin-dependent protein kinase 2 (1:1000, CDK2, Servicebio, Cat No: GB112129), and cyclin B1 (1:1000, Servicebio, Cat No: GB112098) at 4 °C overnight; and then incubated with HRP-conjugated secondary antibody (1:5000) for 30 min at 25 °C.

Techniques: Reverse Transcription Polymerase Chain Reaction, Western Blot

The comparisons of expression levels of miR-4516, Wnt 8B, FZD2, DVL3, FOSL1, and other key genes associated with cell cycle and apoptosis in primary LUAD tumor tissue with those in normal lung tissue using data from the TCGA and GTEx databases in the human samples. ( A ). miR-4516 was down-regulated in the primary LUAD tumor tissue compared to normal lung tissue in the human samples ( p < 0.05). ( B , D – I ). The expressions of CCNA1, Wnt 8B, DVL3, CDK1, CDK2, and CCNB1 were up-regulated in the primary LUAD tumor tissue compared to normal lung tissue in the human samples ( p < 0.01; p < 0.001). ( C ). Overall survival was inversely associated with FOSL1 expressions in patients with LUAD ( p < 0.001). (* p < 0.01; ** p < 0.01; *** p < 0.001).

Journal: Molecules

Article Title: Temporin-GHaK Exhibits Antineoplastic Activity against Human Lung Adenocarcinoma by Inhibiting the Wnt Signaling Pathway through miRNA-4516

doi: 10.3390/molecules29122797

Figure Lengend Snippet: The comparisons of expression levels of miR-4516, Wnt 8B, FZD2, DVL3, FOSL1, and other key genes associated with cell cycle and apoptosis in primary LUAD tumor tissue with those in normal lung tissue using data from the TCGA and GTEx databases in the human samples. ( A ). miR-4516 was down-regulated in the primary LUAD tumor tissue compared to normal lung tissue in the human samples ( p < 0.05). ( B , D – I ). The expressions of CCNA1, Wnt 8B, DVL3, CDK1, CDK2, and CCNB1 were up-regulated in the primary LUAD tumor tissue compared to normal lung tissue in the human samples ( p < 0.01; p < 0.001). ( C ). Overall survival was inversely associated with FOSL1 expressions in patients with LUAD ( p < 0.001). (* p < 0.01; ** p < 0.01; *** p < 0.001).

Article Snippet: Polyvinylidene difluoride membranes (Servicebio, Cat No: G6015-0.45) were blocked with 5% non-fat milk in TBST buffer; incubated with primary antibodies against GAPDH (1:2000, Servicebio, Cat No: GB15004), ACTIN (1:2000, Servicebio, Cat No: GB15003), p-GSK-3β (1:1000, Servicebio, Cat No: GB114582), cyclin A1 (1:500, ABCLONAL, Cat No: A14529), cyclin-dependent protein kinase 1 (1:1000, CDK1, Servicebio, Cat No: GB11398), cyclin-dependent protein kinase 2 (1:1000, CDK2, Servicebio, Cat No: GB112129), and cyclin B1 (1:1000, Servicebio, Cat No: GB112098) at 4 °C overnight; and then incubated with HRP-conjugated secondary antibody (1:5000) for 30 min at 25 °C.

Techniques: Expressing