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    TargetMol lenalidomide selleck cat
    Lenalidomide Selleck Cat, supplied by TargetMol, used in various techniques. Bioz Stars score: 93/100, based on 9 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Average 93 stars, based on 9 article reviews
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    Figure 5. E3 ubiquitin ligase is responsible for CSN5 ubiquitination (A) Western blots representing CSN5, CUL1, CUL2, CUL3, CUL4A, CUL4B, and CUL5 expression levels in MM.1S and RPMI8226 treated with DMSO or 10 mM <t>MLN4924</t> for 12 h. This experiment was performed in triplicate. b-Actin was used as a loading control. Band densities were quantified using iBright Analysis Software. Data are represented as mean ± standard deviation. Student’s t test. (B) Proximity proteomics was performed using RPMI8226 cells overexpressing APEX2-CSN5 or TurboID-CSN5. We identified proteins that were highly enriched (2-fold or more, p < 0.01) in biotin-tyramide- or biotin-treated samples as proximal proteins of CSN5. We selected ubiquitin-related proteins from the enriched proteins and performed an STRING analysis (https://string-db.org/). A protein interaction network generated using STRING is shown. (C) Comparison of the levels of DDB1 and DCN1 expression quantified in proteomics analysis of LEN-sensitive and resistant cell groups among the 15 MM cell lines. Student’s t test. (D) Comparison of the protein levels between CSN5 and DDB1 quantified in proteomics analysis of 15 MM cell lines.
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    Figure 5. E3 ubiquitin ligase is responsible for CSN5 ubiquitination (A) Western blots representing CSN5, CUL1, CUL2, CUL3, CUL4A, CUL4B, and CUL5 expression levels in MM.1S and RPMI8226 treated with DMSO or 10 mM <t>MLN4924</t> for 12 h. This experiment was performed in triplicate. b-Actin was used as a loading control. Band densities were quantified using iBright Analysis Software. Data are represented as mean ± standard deviation. Student’s t test. (B) Proximity proteomics was performed using RPMI8226 cells overexpressing APEX2-CSN5 or TurboID-CSN5. We identified proteins that were highly enriched (2-fold or more, p < 0.01) in biotin-tyramide- or biotin-treated samples as proximal proteins of CSN5. We selected ubiquitin-related proteins from the enriched proteins and performed an STRING analysis (https://string-db.org/). A protein interaction network generated using STRING is shown. (C) Comparison of the levels of DDB1 and DCN1 expression quantified in proteomics analysis of LEN-sensitive and resistant cell groups among the 15 MM cell lines. Student’s t test. (D) Comparison of the protein levels between CSN5 and DDB1 quantified in proteomics analysis of 15 MM cell lines.
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    Figure 5. E3 ubiquitin ligase is responsible for CSN5 ubiquitination (A) Western blots representing CSN5, CUL1, CUL2, CUL3, CUL4A, CUL4B, and CUL5 expression levels in MM.1S and RPMI8226 treated with DMSO or 10 mM <t>MLN4924</t> for 12 h. This experiment was performed in triplicate. b-Actin was used as a loading control. Band densities were quantified using iBright Analysis Software. Data are represented as mean ± standard deviation. Student’s t test. (B) Proximity proteomics was performed using RPMI8226 cells overexpressing APEX2-CSN5 or TurboID-CSN5. We identified proteins that were highly enriched (2-fold or more, p < 0.01) in biotin-tyramide- or biotin-treated samples as proximal proteins of CSN5. We selected ubiquitin-related proteins from the enriched proteins and performed an STRING analysis (https://string-db.org/). A protein interaction network generated using STRING is shown. (C) Comparison of the levels of DDB1 and DCN1 expression quantified in proteomics analysis of LEN-sensitive and resistant cell groups among the 15 MM cell lines. Student’s t test. (D) Comparison of the protein levels between CSN5 and DDB1 quantified in proteomics analysis of 15 MM cell lines.
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    Figure 5. E3 ubiquitin ligase is responsible for CSN5 ubiquitination (A) Western blots representing CSN5, CUL1, CUL2, CUL3, CUL4A, CUL4B, and CUL5 expression levels in MM.1S and RPMI8226 treated with DMSO or 10 mM MLN4924 for 12 h. This experiment was performed in triplicate. b-Actin was used as a loading control. Band densities were quantified using iBright Analysis Software. Data are represented as mean ± standard deviation. Student’s t test. (B) Proximity proteomics was performed using RPMI8226 cells overexpressing APEX2-CSN5 or TurboID-CSN5. We identified proteins that were highly enriched (2-fold or more, p < 0.01) in biotin-tyramide- or biotin-treated samples as proximal proteins of CSN5. We selected ubiquitin-related proteins from the enriched proteins and performed an STRING analysis (https://string-db.org/). A protein interaction network generated using STRING is shown. (C) Comparison of the levels of DDB1 and DCN1 expression quantified in proteomics analysis of LEN-sensitive and resistant cell groups among the 15 MM cell lines. Student’s t test. (D) Comparison of the protein levels between CSN5 and DDB1 quantified in proteomics analysis of 15 MM cell lines.

    Journal: iScience

    Article Title: Increased CSN5 expression enhances the sensitivity to lenalidomide in multiple myeloma cells

    doi: 10.1016/j.isci.2024.111399

    Figure Lengend Snippet: Figure 5. E3 ubiquitin ligase is responsible for CSN5 ubiquitination (A) Western blots representing CSN5, CUL1, CUL2, CUL3, CUL4A, CUL4B, and CUL5 expression levels in MM.1S and RPMI8226 treated with DMSO or 10 mM MLN4924 for 12 h. This experiment was performed in triplicate. b-Actin was used as a loading control. Band densities were quantified using iBright Analysis Software. Data are represented as mean ± standard deviation. Student’s t test. (B) Proximity proteomics was performed using RPMI8226 cells overexpressing APEX2-CSN5 or TurboID-CSN5. We identified proteins that were highly enriched (2-fold or more, p < 0.01) in biotin-tyramide- or biotin-treated samples as proximal proteins of CSN5. We selected ubiquitin-related proteins from the enriched proteins and performed an STRING analysis (https://string-db.org/). A protein interaction network generated using STRING is shown. (C) Comparison of the levels of DDB1 and DCN1 expression quantified in proteomics analysis of LEN-sensitive and resistant cell groups among the 15 MM cell lines. Student’s t test. (D) Comparison of the protein levels between CSN5 and DDB1 quantified in proteomics analysis of 15 MM cell lines.

    Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Anti-Ikaros (IKZF1) Antibody Cell Signaling Technology Cat#5443; RRID: AB_10691693 Anti-Aiolos (IKZF3) Antibody, clone 9D10 Millipore Cat#MABE911 Anti-rabbit IgG, HRP-linked Antibody Cell Signaling Technology Cat#7074; RRID: AB_2099233 Anti-b-Actin pAb-HRP-DirecT MBL Life Science Cat#PM053-7; RRID: AB_10697035 CSN5 Rabbit pAb Cell Signaling Technology Cat#6895; RRID: AB_10839271 CUL1 Rabbit pAb Cell Signaling Technology Cat#4995; RRID: AB_2261133 CUL2 (C-4) Mouse mAb Santa Cruz Biotechnology Cat#sc-166506; RRID: AB_2230072 CUL3 Rabbit pAb Cell Signaling Technology Cat#2759; RRID: AB_2086432 CUL4A Rabbit pAb abcam Cat#ab72548; RRID: AB_1268363 CUL4B Rabbit pAb Proteintech Cat#12916-1-AP; RRID: AB_2086699 CUL5 (F-6) Mouse mAb Santa Cruz Biotechnology Cat#sc-373822; RRID: AB_10992228 FLAG Mouse mAb, M2 SIGMA Cat#F3165; RRID: AB_259529 Goat anti-Mouse IgG (H + L) Secondary Antibody, HRP Thermo Fisher Scientific Cat#62–6520; RRID: AB_88369 IKKa Rabbit pAb (H-744) Santa Cruz Biotechnology Cat#sc-7218; RRID: AB_2079399 IKKb Rabbit pAb Cell Signaling Technology Cat#2684; RRID: AB_2122298 IRF4 Rabbit pAb Cell Signaling Technology Cat#4964; RRID: AB_10698467 LC3 Mouse mAb MBL Life Science Cat#M152-3; RRID: AB_1279144 Phospho-IKKa (Ser176)/IKKb (Ser177) (C84E11) Rabbit mAb Cell Signaling Technology Cat#2078; RRID: AB_2079379 Phospho-IKKa/b (Ser176/180) (16A6) Rabbit mAb Cell Signaling Technology Cat#2697; RRID: AB_2079382 Ubiquitin (P4D1) Mouse mAb Santa Cruz Biotechnology Cat#sc-8017; RRID: AB_628423 Chemicals, peptides, and recombinant proteins 2-chloroacetamide Wako Cat#032-09762 Dithiothreitol Wako Cat#049-08972 MG132 Selleck Cat#S2619 MLN4924 Selleck Cat#S7109 Lenalidomide Selleck Cat#S1029 Bafilomycin A1 Selleck Cat#S1413 Cycloheximide Wako Cat#035-20992 Lysyl Endopeptidase Wako Cat#121-05063 Sequencing Grade Modified Trypsin Promega Cat#V5113 Sodium deoxycholate Wako Cat#194-08311 Sodium lauroyl sarcosinate Wako Cat#196-10385 Critical commercial assays BCA Protein Assay Kit Thermo Fisher Scientific Cat #23227 PTMSscan HS K-e-GG Remnant Magnetic Immunoaffinity Beads Cell Signaling Technology Cat#5562 Deposited data Mass spectrometry data This paper jPOST/ProteomeXchange: JPST002950/PXD050020, JPST003373/PXD056174 Original western blot images This paper Mendeley: https://doi.org/10.17632/ggpw7kjjdb.1 Proteomics data of 15 MM cell lines This paper MM Proteome Data: https://mmproteomicsdata.iab.keio.ac.jp/ (Continued on next page) e1 iScience 27, 111399, December 20, 2024

    Techniques: Ubiquitin Proteomics, Western Blot, Expressing, Control, Software, Standard Deviation, Generated, Comparison