plasmid pdsp1  (Addgene inc)

Journal: The EMBO Journal

Article Title: SDCBP/Syntenin-1 stabilizes BACH1 by disassembling the SCF FBXO22 –BACH1 complex in triple-negative breast cancer

doi: 10.1038/s44318-025-00440-1

Figure Lengend Snippet: ( A ) Crosslink immunoprecipitation showing an interaction of FBXO22 with SDCBP in Hs578T cells transfected with Myc-SDCBP-expressing vector or Myc-SDCBP_Δ4-expressing vector. Schematic showing the FBXO22 mutant constructs generated using the HA-FBXO22 plasmid. ( C ) Immunoprecipitation showing SDCBP interactions with FBXO22 and its mutant constructs in HEK293 cells transfected with the indicated plasmids. ns indicates non-specific bands. See also Fig. , . ( D ) His-Pulldown assay showing the effect of SDCBP on SKP1-CUL1-FBXO22 complex formation after the indicated transfections in HEK293 cells. See also Appendix Fig. S . ( E ) His-Pulldown assay showing the effect of SDCBP KO on the SCF FBXO22 –BACH1 complex formation in the scramble control and SDCBP-KO MDA-MB-231 cells transfected with control vector or His-SKP1-expressing vector. See also Appendix Fig. S . ( F ) Immunoprecipitation showing the effect of SDCBP KD on the SCF FBXO22 –BACH1 complex formation in MDA-MB-231 cells transfected with scramble or SDCBP siRNA. ( G ) Immunoprecipitation showing the effect of SDCBP overexpression on the SCF FBXO22 –BACH1 complex formation in Hs578T cells transfected with control vector or Myc-SDCBP-overexpressing vector. ( H ) Immunoprecipitation showing the effect of SDCBP PDZ1 domain on the SCF FBXO22 –BACH1 complex formation in Hs578T cells transfected with a control vector or a Myc-SDCBP-PDZ1-overexpressing vector. .

Article Snippet: pCMV3-N-FLAG-CUL1 cDNA , Sino Biological , Cat#HG17691-NF.

Techniques: Immunoprecipitation, Transfection, Expressing, Plasmid Preparation, Mutagenesis, Construct, Generated, Control, Over Expression

Journal: Nature communications

Article Title: Chemoproteogenomic stratification of the missense variant cysteinome.

doi: 10.1038/s41467-024-53520-x

Figure Lengend Snippet: Fig. 9 | Functional studies of CAND1 and HMGB1. A WT and G1069C mutant CAND1 proteins bind Cul1 while the G1069C CAND1 mutation perturbs binding. HEK293T cells were co-transfected with FLAG-Cul1 and the given HA-tagged CAND1 protein (WT, G1069C, or G1069W) or control FLAG-GFP. Anti-FLAG resin was used to pull down FLAG-Cul1 from cell lysates along with any complexed proteins. Western Blots were incubated with the indicated primary antibodies, *indicates a non-specific HA band. B HMGB1 proteins were tested for the ability to induce TLR4-mediated immune response using HEK-Blue reporter cell lines (hTLR4 and Null control) and corresponding PRR assay. Results show mean response ratios (error bars = SD, n = 4 per condition) of hTLR4 and Null cells to increasing concentrations (μg/mL) of WT, R110C, and R110W proteins as indicated over 2 independent experiments. AT = commercially available all-thiol fully

Article Snippet: HEK293T cells were co-transfected with 3xFLAG-CUL1-pCMV7.1 (Addgene, plasmid #155019) alongwith the given CAND1 plasmid (WT, G1069C, G1069W) or control FLAG‐GFP plasmid.

Techniques: Functional Assay, Mutagenesis, Binding Assay, Transfection, Control, Western Blot, Incubation