shRNA:Article Title: The Crosstalk Between CRL5 and APC/C E3 Ligases Regulates Metastasis and Chemosensitivity of Cancer Cells.
Article Snippet: .. [63] The following antibodies were used: ACTIN (Sigma–Aldrich, A5441), FLAG (Sigma–Aldrich, F1804), FLAG (Sigma–Aldrich, F7425), GFP (Santa Cruz, sc-9996), MYC (Santa Cruz, sc-789), APC2 (Cell Signaling Technology, 12301), APC8 (Cell Signaling Technology, 15100), APC10 (Cell Signaling Technology, 14807), APC11 (Cell Signaling Technology, 14090), APC11 (Santa Cruz, sc-517142), SAG (Proteintech, 11905-1-AP), RBX1 (Proteintech, 14895-1-AP), CUL1 (Santa Cruz, sc-11384), CUL5 (Abcam, ab184177), CUL5 (Santa Cruz, sc-373822), CAND1 (Abcam, ab183748), integrin β1 (Cell Signaling Technology, 34 971), p-SRC (Y416) (Cell Signaling Technology, 6943), ATM (Cell Signaling Technology, 2873), Cyclin B1 (Cell Signaling Technology, 4138), RPB1 (Cell Signaling Technology, 14958), EGFR (Cell Signaling Technology, 4267), DEPTOR (Cell Signaling Technology, 11 816), NOXA (Millipore, OP180), UBE2F (Proteintech, 17056-1-AP), COPS5 (Cell Signaling Technology, 6895), SOCS3 (Proteintech, 14025-1-AP), LC3 (Cell Signaling Technology, 2775), securin (Cell Signaling Technology, 13445), ITCH (Proteintech, 20920-1-AP), Cyclin E1 (Cell Signaling Technology, 20808), p-H3 (Cell Signaling Technology, 3377), and p-Aurora A/B/C (Cell Signaling Technology, 2914), Aurora A (Cell Signaling Technology, 14 475), Aurora B (Abcam, ab2254), and NEDD8 (Abcam, ab81264). siRNA and Lentiviral shRNA Silencing: Cells were transfected with the following siRNA oligos by Lipofectamine 2000 (Invitrogen, 11668019) to achieve transient knockdown of endogenous genes. siCtrl: 5′-ATT GTA TGC GAT CGC AGA C-3′; siCUL5: 5′-GCT AGA ATG TTT CAG GAC ATA-3′; siCUL5-2: 5′-GTC TCA CTT CCT ACT GAA CTG-3′; siAPC11: 5′-TCT GCA GGA TGG CAT TTA A-3′; siAPC11-2: 5′-CCA CAT GCA TTG CAT CCT CAA3′; siAPC11-3: 5′-ACT CAT TAA ACT ACT CAA ATC-3′; siAPC2: 5′-TGC GCG GAGTCT TGT TCT TTA-3′; siAPC2-2: 5′-GCAGAT TAA AGCAAG TCAGAT3′; siSAG: 5′-CCT GTG GGT GAA ACA GAA CAA-3′; siSAG-2: 5′-CGA CAA GAT GTT CTC CCT CAA-3′; siUBE2F: 5′-CAA AGT GAA ATG CCT GAC CAA-3′; siUBE2F-2: 5′-CAT CAA ACG TTA TGC CAG AT-3′; siintegrin β1: 5′-GCC CTC CAG ATG ACA TAG AAA-3′; siintegrin β1-2: 5′-GCC TTG CAT TAC TGC TGA TAT-3′; siITCH: 5′-GCC TAT GTT CGG GAC TTC AAA-3′; siITCH-2: 5′-GGT GAC AAA GAG CCA ACA GAG-3′; siWSB1: 5′-AGT TTC TCT CGT ATC GTA TTT-3′; siAREL1: 5′-CCG GGA ATG GTT TGA GCT AAT3′; siTRIP12: 5′-TCG CAA AGG TTA AGA TGA A-3′; siTRAF4: 5′-CCA GGA CAT TCG AAA GCG AAA-3′; siCOPS5: 5′-GCT CAG AGT ATC GAT GAA A-3′; and siCOPS5-2: 5′-CAG TCT CTG AGA AGT ACT TTA-3′. ..
Transfection:Article Title: The Crosstalk Between CRL5 and APC/C E3 Ligases Regulates Metastasis and Chemosensitivity of Cancer Cells.
Article Snippet: .. [63] The following antibodies were used: ACTIN (Sigma–Aldrich, A5441), FLAG (Sigma–Aldrich, F1804), FLAG (Sigma–Aldrich, F7425), GFP (Santa Cruz, sc-9996), MYC (Santa Cruz, sc-789), APC2 (Cell Signaling Technology, 12301), APC8 (Cell Signaling Technology, 15100), APC10 (Cell Signaling Technology, 14807), APC11 (Cell Signaling Technology, 14090), APC11 (Santa Cruz, sc-517142), SAG (Proteintech, 11905-1-AP), RBX1 (Proteintech, 14895-1-AP), CUL1 (Santa Cruz, sc-11384), CUL5 (Abcam, ab184177), CUL5 (Santa Cruz, sc-373822), CAND1 (Abcam, ab183748), integrin β1 (Cell Signaling Technology, 34 971), p-SRC (Y416) (Cell Signaling Technology, 6943), ATM (Cell Signaling Technology, 2873), Cyclin B1 (Cell Signaling Technology, 4138), RPB1 (Cell Signaling Technology, 14958), EGFR (Cell Signaling Technology, 4267), DEPTOR (Cell Signaling Technology, 11 816), NOXA (Millipore, OP180), UBE2F (Proteintech, 17056-1-AP), COPS5 (Cell Signaling Technology, 6895), SOCS3 (Proteintech, 14025-1-AP), LC3 (Cell Signaling Technology, 2775), securin (Cell Signaling Technology, 13445), ITCH (Proteintech, 20920-1-AP), Cyclin E1 (Cell Signaling Technology, 20808), p-H3 (Cell Signaling Technology, 3377), and p-Aurora A/B/C (Cell Signaling Technology, 2914), Aurora A (Cell Signaling Technology, 14 475), Aurora B (Abcam, ab2254), and NEDD8 (Abcam, ab81264). siRNA and Lentiviral shRNA Silencing: Cells were transfected with the following siRNA oligos by Lipofectamine 2000 (Invitrogen, 11668019) to achieve transient knockdown of endogenous genes. siCtrl: 5′-ATT GTA TGC GAT CGC AGA C-3′; siCUL5: 5′-GCT AGA ATG TTT CAG GAC ATA-3′; siCUL5-2: 5′-GTC TCA CTT CCT ACT GAA CTG-3′; siAPC11: 5′-TCT GCA GGA TGG CAT TTA A-3′; siAPC11-2: 5′-CCA CAT GCA TTG CAT CCT CAA3′; siAPC11-3: 5′-ACT CAT TAA ACT ACT CAA ATC-3′; siAPC2: 5′-TGC GCG GAGTCT TGT TCT TTA-3′; siAPC2-2: 5′-GCAGAT TAA AGCAAG TCAGAT3′; siSAG: 5′-CCT GTG GGT GAA ACA GAA CAA-3′; siSAG-2: 5′-CGA CAA GAT GTT CTC CCT CAA-3′; siUBE2F: 5′-CAA AGT GAA ATG CCT GAC CAA-3′; siUBE2F-2: 5′-CAT CAA ACG TTA TGC CAG AT-3′; siintegrin β1: 5′-GCC CTC CAG ATG ACA TAG AAA-3′; siintegrin β1-2: 5′-GCC TTG CAT TAC TGC TGA TAT-3′; siITCH: 5′-GCC TAT GTT CGG GAC TTC AAA-3′; siITCH-2: 5′-GGT GAC AAA GAG CCA ACA GAG-3′; siWSB1: 5′-AGT TTC TCT CGT ATC GTA TTT-3′; siAREL1: 5′-CCG GGA ATG GTT TGA GCT AAT3′; siTRIP12: 5′-TCG CAA AGG TTA AGA TGA A-3′; siTRAF4: 5′-CCA GGA CAT TCG AAA GCG AAA-3′; siCOPS5: 5′-GCT CAG AGT ATC GAT GAA A-3′; and siCOPS5-2: 5′-CAG TCT CTG AGA AGT ACT TTA-3′. ..
Knockdown:Article Title: The Crosstalk Between CRL5 and APC/C E3 Ligases Regulates Metastasis and Chemosensitivity of Cancer Cells.
Article Snippet: .. [63] The following antibodies were used: ACTIN (Sigma–Aldrich, A5441), FLAG (Sigma–Aldrich, F1804), FLAG (Sigma–Aldrich, F7425), GFP (Santa Cruz, sc-9996), MYC (Santa Cruz, sc-789), APC2 (Cell Signaling Technology, 12301), APC8 (Cell Signaling Technology, 15100), APC10 (Cell Signaling Technology, 14807), APC11 (Cell Signaling Technology, 14090), APC11 (Santa Cruz, sc-517142), SAG (Proteintech, 11905-1-AP), RBX1 (Proteintech, 14895-1-AP), CUL1 (Santa Cruz, sc-11384), CUL5 (Abcam, ab184177), CUL5 (Santa Cruz, sc-373822), CAND1 (Abcam, ab183748), integrin β1 (Cell Signaling Technology, 34 971), p-SRC (Y416) (Cell Signaling Technology, 6943), ATM (Cell Signaling Technology, 2873), Cyclin B1 (Cell Signaling Technology, 4138), RPB1 (Cell Signaling Technology, 14958), EGFR (Cell Signaling Technology, 4267), DEPTOR (Cell Signaling Technology, 11 816), NOXA (Millipore, OP180), UBE2F (Proteintech, 17056-1-AP), COPS5 (Cell Signaling Technology, 6895), SOCS3 (Proteintech, 14025-1-AP), LC3 (Cell Signaling Technology, 2775), securin (Cell Signaling Technology, 13445), ITCH (Proteintech, 20920-1-AP), Cyclin E1 (Cell Signaling Technology, 20808), p-H3 (Cell Signaling Technology, 3377), and p-Aurora A/B/C (Cell Signaling Technology, 2914), Aurora A (Cell Signaling Technology, 14 475), Aurora B (Abcam, ab2254), and NEDD8 (Abcam, ab81264). siRNA and Lentiviral shRNA Silencing: Cells were transfected with the following siRNA oligos by Lipofectamine 2000 (Invitrogen, 11668019) to achieve transient knockdown of endogenous genes. siCtrl: 5′-ATT GTA TGC GAT CGC AGA C-3′; siCUL5: 5′-GCT AGA ATG TTT CAG GAC ATA-3′; siCUL5-2: 5′-GTC TCA CTT CCT ACT GAA CTG-3′; siAPC11: 5′-TCT GCA GGA TGG CAT TTA A-3′; siAPC11-2: 5′-CCA CAT GCA TTG CAT CCT CAA3′; siAPC11-3: 5′-ACT CAT TAA ACT ACT CAA ATC-3′; siAPC2: 5′-TGC GCG GAGTCT TGT TCT TTA-3′; siAPC2-2: 5′-GCAGAT TAA AGCAAG TCAGAT3′; siSAG: 5′-CCT GTG GGT GAA ACA GAA CAA-3′; siSAG-2: 5′-CGA CAA GAT GTT CTC CCT CAA-3′; siUBE2F: 5′-CAA AGT GAA ATG CCT GAC CAA-3′; siUBE2F-2: 5′-CAT CAA ACG TTA TGC CAG AT-3′; siintegrin β1: 5′-GCC CTC CAG ATG ACA TAG AAA-3′; siintegrin β1-2: 5′-GCC TTG CAT TAC TGC TGA TAT-3′; siITCH: 5′-GCC TAT GTT CGG GAC TTC AAA-3′; siITCH-2: 5′-GGT GAC AAA GAG CCA ACA GAG-3′; siWSB1: 5′-AGT TTC TCT CGT ATC GTA TTT-3′; siAREL1: 5′-CCG GGA ATG GTT TGA GCT AAT3′; siTRIP12: 5′-TCG CAA AGG TTA AGA TGA A-3′; siTRAF4: 5′-CCA GGA CAT TCG AAA GCG AAA-3′; siCOPS5: 5′-GCT CAG AGT ATC GAT GAA A-3′; and siCOPS5-2: 5′-CAG TCT CTG AGA AGT ACT TTA-3′. ..
Western Blot:Article Title: Innate immune sensing of Z-nucleic acids by ZBP1-RIPK1 axis drives neuroinflammation in Alzheimer's disease.
Article Snippet: In brief Microglia-mediated neuroinflammation drives Alzheimer’s disease (AD) pathogenesis, yet the underlying mechanism is poorly understood.. Song et al. demonstrate that toxic amyloid-β induces oxidation and fragmentation of mitochondrial DNA in microglia, facilitating its B-to-Z conformational transition and subsequent recognition by the innate immune sensor ZBP1, which activates RIPK1 kinase and drives detrimental immune responses and neuroinflammation in AD.
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