cullin1  (Addgene inc)

Journal: Cancers

Article Title: Pimozide Reprograms the Ran GTPase–SCF Axis and Matrix Remodeling Pathways in Breast, Colorectal, and Pancreatic Cancer Models

doi: 10.3390/cancers18040611

Figure Lengend Snippet: Gene expression levels of ( a ) Ran , ( b ) MMP2 , ( c ) FBXW10 , and ( d ) Cullin1 in MCF7 cell line after treatment with PMZ. An unpaired t -test was used to analyze qPCR results for the MDA-MB-231 cell line after treatment with PMZ (IC 50 = 16.02 µM) for 48 h in triplicate. The housekeeping gene β2M was used for normalizing the data. *** p < 0.001, ns = not significant.

Article Snippet: Trypsin and primers, including FBXW10 (F: 5′-AACAGCACCCAGTGGACCAA-3′/R: 5′-TGTCTTGATTGAGCCCTGAGAT-3′), β2M (F: 5′-GAGGCTATCCAGCGTACTCCA-3′/R: 5′-CGGCAGGCATACTCATCTTTT-3′), Rbx1 E3 Ligase (F: 5′-ATGCCCCAATCTTGTCCATCT-3′/R: 5′-CACCGACTGAGTGATAGGTGT-3′), Skp2 (F: 5′-ATGCCCCAATCTTGTCCATCT-3′/R: 5′-CACCGACTGAGTGATAGGTGT-3′), MMP2 (F: 5′-TACAGGATCATTGGCTACACACC-3′/R: 5′-GGTCACATCGCTCCAGACR-3′), RAN (F: 5′-TCTGGCTTGCTAGGAAGCTCA-3′/R: 5′-GCTGGGTCCATGACAACTTCT-3′), and Cullin1 (F: 5′-AGCCATTGAAAAGTGGAGAA-3′/R: 5′-GCGTCATTGTTGAATGCAGACA-3′), were purchased from Macrogen Humanizing Genomics (Seoul, Republic of Korea).

Techniques: Gene Expression

Journal: Cancers

Article Title: Pimozide Reprograms the Ran GTPase–SCF Axis and Matrix Remodeling Pathways in Breast, Colorectal, and Pancreatic Cancer Models

doi: 10.3390/cancers18040611

Figure Lengend Snippet: Gene expression levels of ( a ) Ran, ( b ) MMP2, ( c ) FBXW10, ( d ) Cullin1, and ( e ) Rbx1 in HT-29 cell line after treatment with PMZ. An unpaired t -test was used to analyze qPCR results for the MDA-MB-231 cell line after treatment with PMZ (IC 50 = 35.41 µM) for 48 h in triplicate. The housekeeping gene β2M was used for normalizing the data. * p < 0.05, *** p < 0.001.

Article Snippet: Trypsin and primers, including FBXW10 (F: 5′-AACAGCACCCAGTGGACCAA-3′/R: 5′-TGTCTTGATTGAGCCCTGAGAT-3′), β2M (F: 5′-GAGGCTATCCAGCGTACTCCA-3′/R: 5′-CGGCAGGCATACTCATCTTTT-3′), Rbx1 E3 Ligase (F: 5′-ATGCCCCAATCTTGTCCATCT-3′/R: 5′-CACCGACTGAGTGATAGGTGT-3′), Skp2 (F: 5′-ATGCCCCAATCTTGTCCATCT-3′/R: 5′-CACCGACTGAGTGATAGGTGT-3′), MMP2 (F: 5′-TACAGGATCATTGGCTACACACC-3′/R: 5′-GGTCACATCGCTCCAGACR-3′), RAN (F: 5′-TCTGGCTTGCTAGGAAGCTCA-3′/R: 5′-GCTGGGTCCATGACAACTTCT-3′), and Cullin1 (F: 5′-AGCCATTGAAAAGTGGAGAA-3′/R: 5′-GCGTCATTGTTGAATGCAGACA-3′), were purchased from Macrogen Humanizing Genomics (Seoul, Republic of Korea).

Techniques: Gene Expression

Journal: Inflammation

Article Title: Isoorientin Ameliorates Macrophage Pyroptosis and Atherogenesis by Reducing KDM4A Levels and Promoting SKP1-Cullin1-F-box E3 Ligase-mediated NLRP3 Ubiquitination

doi: 10.1007/s10753-025-02289-2

Figure Lengend Snippet: SKP1 reduces the assembly and activation of the NLRP3 inflammasome through ubiquitination. A , B , Colocalization of NLRP3 with SKP1 ( A ) and Cullin1 ( B ) in BMDMs determined using double-label immunofluorescence staining ( n = 3). C , D , Binding relationships between NLRP3 and SKP1 ( C ) and between NLRP3 and Cullin1 ( D ) in BMDMs determined via co-IP assays ( n = 3). E , Effects of SKP1 on NLRP3 ubiquitination determined via co-IP assays upon transfection of BMDMs with Flag-SKP1, HA-NLRP3, or Myc-Ub ( n = 3). F , Interactions between NLRP3 and SKP1 and ASCs in BMDMs overexpressing SKP1 determined via co-IP assays ( n = 3). BMDMs were transfected with oe-KDM4A or oe-SKP1, followed by ox-LDL induction and ISO treatment. G , SKP1 protein levels in BMDMs after transfection determined using WB analysis ( n = 3); H Positive staining of ASCs in transfected BMDMs detected using immunofluorescence staining ( n = 3). Differences were compared by the unpaired t test or one-way ANOVA followed by Tukey’s post hoc test. Each dot represents an independent experiment. * p < 0.05

Article Snippet: The sections were incubated with antibodies against F4/80 (1:50, ab6640, Abcam, Inc., Cambridge, MA, USA), N-GSDMD (1:100, #DF13758, Affinity Biosciences, OH, USA), cleaved caspase 1 (1:50, HY- P80622 , MedChemExpress, Monmouth Junction, NJ, USA), KDM4A (1:100, ab191433, Abcam), SKP1 (1:500, 67,745–1-Ig, Proteintech Group, Inc., Wuhan, Hubei, China), NLRP3 (1:100, 30,109–1-AP, Proteintech), Cullin1 (1:200, 66,978–1-Ig, Proteintech), and ASC (1:100, sc-514414, Santa Cruz Biotechnology, Inc., Santa Cruz, CA, USA).

Techniques: Activation Assay, Ubiquitin Proteomics, Immunofluorescence, Staining, Binding Assay, Co-Immunoprecipitation Assay, Transfection

Journal: Cells

Article Title: CCT and Cullin1 Regulate the TORC1 Pathway to Promote Dendritic Arborization in Health and Disease

doi: 10.3390/cells13121029

Figure Lengend Snippet: CCT and the TORC1 pathway promote dendritic arborization . ( A ) Schematic diagram of regulatory relationships between the insulin pathway, SCF complex, CCT, and TORC1 pathway, with the insulin pathway indicated by teal arrows. TORC1 is negatively regulated by Cullin1 and positively regulated by CCT. The upstream insulin pathway in green is displayed for context, but was not examined in this study. Individual components of the SCF and TORC1 complexes examined in this study are outlined in white. ( B ) Representative images of CIV neurons for key CCT and TORC1 pathway manipulations, with RNAi-mediated knockdown indicated with -IR and UAS-mediated overexpression with -OE. Scale bars = 100 µm ( C ) Total dendritic length of CCT and TORC1 pathway manipulations shown in comparison to a WT control. ( D ) Number of Sholl maximum intersections. ( E ) Radius (in µm) of Sholl maximum intersection for each genotype. Radii that have shifted a significant difference from control are indicated with an asterisk. In all panels * = p < 0.05, see for detailed statistics.

Article Snippet: The primary antibodies used included: chicken anti-GFP (1:1000 dilution, Aves Labs, Davis, CA, USA), rabbit anti-phosphorylated S6k (1:300 dilution, Cell Signaling Technology, Danvers, MA, USA), rabbit anti-Raptor (1:200 dilution, Cell Signaling Technology, Danvers, MA, USA), mouse anti-acetylated α-tubulin (1:100 dilution, Santa Cruz Biotechnology, Dallas, TX, USA), mouse anti-Futsch (1:100 dilution, Developmental Studies Hybridoma Bank, Iowa City, IA, USA), rabbit anti-Huntingtin (1:200 dilution, Cell Signaling Technology, Danvers, MA, USA), mouse anti-CCT5 (1:200 dilution, GeneTex, Irvine, CA, USA), mouse anti-S6k (1:200 dilution, Proteintech, Rosemont, IL, USA), rabbit anti-phosphorylated Akt (1:200 dilution, Cell Signaling Technology, Danvers, MA, USA), rabbit anti-Cullin1 (1:200 dilution, Invitrogen), and mouse anti-β-tubulin IIA (1:500 dilution, Novus Biologicals, Centennial, CO, USA).

Techniques: Knockdown, Over Expression, Comparison, Control

Journal: Cells

Article Title: CCT and Cullin1 Regulate the TORC1 Pathway to Promote Dendritic Arborization in Health and Disease

doi: 10.3390/cells13121029

Figure Lengend Snippet: CCT and Cullin1 regulate the TORC1 pathway in vivo . ( A ) Heat map showing the percent change in P-S6k fluorescence for each genetic manipulation as compared to its proper control. See for representative images. ( B ) Raptor fluorescence is significantly decreased in CCT5 LOF conditions and is not rescued by overexpression of Raptor. ( C ) P-S6k fluorescence levels are significantly decreased in CCT5 LOF and are not rescued by overexpression of S6k. ( D ) Representative images of combined TORC1 genetic manipulations. Scale bars = 100 µm. ( E ) Total dendritic length in microns for WT and combined TORC1 genetic manipulations. ( F ) Number of Sholl maximum intersections for S6k and Cullin1 individual and combined LOF. In all panels * = p < 0.05, ns = not significant; see for detailed statistics.

Article Snippet: The primary antibodies used included: chicken anti-GFP (1:1000 dilution, Aves Labs, Davis, CA, USA), rabbit anti-phosphorylated S6k (1:300 dilution, Cell Signaling Technology, Danvers, MA, USA), rabbit anti-Raptor (1:200 dilution, Cell Signaling Technology, Danvers, MA, USA), mouse anti-acetylated α-tubulin (1:100 dilution, Santa Cruz Biotechnology, Dallas, TX, USA), mouse anti-Futsch (1:100 dilution, Developmental Studies Hybridoma Bank, Iowa City, IA, USA), rabbit anti-Huntingtin (1:200 dilution, Cell Signaling Technology, Danvers, MA, USA), mouse anti-CCT5 (1:200 dilution, GeneTex, Irvine, CA, USA), mouse anti-S6k (1:200 dilution, Proteintech, Rosemont, IL, USA), rabbit anti-phosphorylated Akt (1:200 dilution, Cell Signaling Technology, Danvers, MA, USA), rabbit anti-Cullin1 (1:200 dilution, Invitrogen), and mouse anti-β-tubulin IIA (1:500 dilution, Novus Biologicals, Centennial, CO, USA).

Techniques: In Vivo, Fluorescence, Control, Over Expression

Journal: Cells

Article Title: CCT and Cullin1 Regulate the TORC1 Pathway to Promote Dendritic Arborization in Health and Disease

doi: 10.3390/cells13121029

Figure Lengend Snippet: Expression of mHTT leads to dendritic hypotrophy parallel to TORC1 pathway . ( A ) Representative image of WT HTT staining in CIV neuron—dendrite marked by dashed white lines, axon by solid lines. Scale bar = 3 µm. ( B ) Representative images of mHTT25-Cerulean and mHTT96-Cerulean shown with aggregate inclusion bodies marked by white arrows for mHTT96-Cerulean. Scale bar = 10 µm. ( C ) Representative reconstructions of branches from WT and TORC1 genetic manipulations—normalized mCherry::Jupiter fluorescence is coded with the rainbow spectrum shown (A.U.) ( D ) Heat map representing the average normalized, binned mCherry::Jupiter fluorescence along dendrites at increasing distances from the soma for overexpressions of mHTT 20, 50, and 93 repeats. Genotypes found to be significantly different along the dendritic arbor are marked with an asterisk. ( E ) TDL of Cullin1-IR and CCT5-IR in both mHTTQ25 and mHTTQ96 backgrounds displayed as percent change from WT control. CCT5-IR decreases both mHTTQ96 and mHTTQ25 neurons to far lower than WT, while Cullin1-IR rescues mHTT96 hypotrophy to WT levels. CCT5-IR alone is not significantly different from either mHTTQ25;CCT5-IR or mHTTQ96;CCT5-IR. All genotypes vary significantly from WT except for mHTTQ96;Cullin1-IR. ( F ) Representative images of CIV dendritic morphology in combined HTT and CCT5-IR or Cullin1-IR combinations. Scale bars = 100 µm. ( G ) Number of mHTT aggregate IBs does not change due to CCT5 or Cullin1 LOF. ( H ) mHTT aggregates in mHTTQ96Cerulean conditions do not change in average area due to CCT5 or Cullin1 LOF. In all panels * = p < 0.05, ns = not significant; see for detailed statistics.

Article Snippet: The primary antibodies used included: chicken anti-GFP (1:1000 dilution, Aves Labs, Davis, CA, USA), rabbit anti-phosphorylated S6k (1:300 dilution, Cell Signaling Technology, Danvers, MA, USA), rabbit anti-Raptor (1:200 dilution, Cell Signaling Technology, Danvers, MA, USA), mouse anti-acetylated α-tubulin (1:100 dilution, Santa Cruz Biotechnology, Dallas, TX, USA), mouse anti-Futsch (1:100 dilution, Developmental Studies Hybridoma Bank, Iowa City, IA, USA), rabbit anti-Huntingtin (1:200 dilution, Cell Signaling Technology, Danvers, MA, USA), mouse anti-CCT5 (1:200 dilution, GeneTex, Irvine, CA, USA), mouse anti-S6k (1:200 dilution, Proteintech, Rosemont, IL, USA), rabbit anti-phosphorylated Akt (1:200 dilution, Cell Signaling Technology, Danvers, MA, USA), rabbit anti-Cullin1 (1:200 dilution, Invitrogen), and mouse anti-β-tubulin IIA (1:500 dilution, Novus Biologicals, Centennial, CO, USA).

Techniques: Expressing, Staining, Fluorescence, Control

Journal: iScience

Article Title: The HRD1-SEL1L ubiquitin ligase regulates stress granule homeostasis in couple with distinctive signaling branches of ER stress

doi: 10.1016/j.isci.2024.110196

Figure Lengend Snippet:

Article Snippet: Rabbit anti-Cullin1 , Invitrogen , Cat# 71-8700; RRID: AB_2534002.

Techniques: Recombinant, Bicinchoninic Acid Protein Assay, Software