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nature communications  (ATCC)
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nature communications  (Addgene inc)
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cand1 cul1 sr cand2 cul1 nature communications  (Santa Cruz Biotechnology)
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Santa Cruz Biotechnology cand1 cul1 sr cand2 cul1 nature communications
Fig. 1 | <t>CAND2</t> promotes SCF-mediated protein degradation. A CAND2 binds to unneddylated <t>CUL1</t> in human cell lysates. Whole-cell lysates (WCL) from HEK293 cells of the indicated genotypes were immunoprecipitated (IP) with an anti-HA antibody. Cells were treated with 1 μM MLN4924 for 1 h before harvesting. Samples were immunoblotted with anti-CUL1, anti-HA, and anti-GAPDH antibodies. DKO: <t>CAND1</t> and CAND2 double knockout cells; DKOCAND1-HA: DKO cells expressing transgenic CAND1HA; DKOCAND2-HA: DKO cells expressing transgenic CAND2HA. Representative results from three independent experiments are shown. B, C CAND2 rescues the TNFα-induced degradation of p-IκBα in DKO cells. B Immunoblotting of HA (CAND1 or CAND2), IκBα, and GAPDH at the indicated time points after the addition of TNFα to the indicated cells in the presence of cycloheximide (CHX). C Quantification of the relative p-IκBα levels in (B). Data are presented as mean ± SEM; n = 3 independent experiments. D, E Both CAND1 and CAND2 are required for proper degradation of IRP2 in HEK293 cells.
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k nature communications  (Addgene inc)
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Addgene inc k nature communications
Fig. 1 | <t>CAND2</t> promotes SCF-mediated protein degradation. A CAND2 binds to unneddylated <t>CUL1</t> in human cell lysates. Whole-cell lysates (WCL) from HEK293 cells of the indicated genotypes were immunoprecipitated (IP) with an anti-HA antibody. Cells were treated with 1 μM MLN4924 for 1 h before harvesting. Samples were immunoblotted with anti-CUL1, anti-HA, and anti-GAPDH antibodies. DKO: <t>CAND1</t> and CAND2 double knockout cells; DKOCAND1-HA: DKO cells expressing transgenic CAND1HA; DKOCAND2-HA: DKO cells expressing transgenic CAND2HA. Representative results from three independent experiments are shown. B, C CAND2 rescues the TNFα-induced degradation of p-IκBα in DKO cells. B Immunoblotting of HA (CAND1 or CAND2), IκBα, and GAPDH at the indicated time points after the addition of TNFα to the indicated cells in the presence of cycloheximide (CHX). C Quantification of the relative p-IκBα levels in (B). Data are presented as mean ± SEM; n = 3 independent experiments. D, E Both CAND1 and CAND2 are required for proper degradation of IRP2 in HEK293 cells.
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Image Search Results


Fig. 1 | CAND2 promotes SCF-mediated protein degradation. A CAND2 binds to unneddylated CUL1 in human cell lysates. Whole-cell lysates (WCL) from HEK293 cells of the indicated genotypes were immunoprecipitated (IP) with an anti-HA antibody. Cells were treated with 1 μM MLN4924 for 1 h before harvesting. Samples were immunoblotted with anti-CUL1, anti-HA, and anti-GAPDH antibodies. DKO: CAND1 and CAND2 double knockout cells; DKOCAND1-HA: DKO cells expressing transgenic CAND1HA; DKOCAND2-HA: DKO cells expressing transgenic CAND2HA. Representative results from three independent experiments are shown. B, C CAND2 rescues the TNFα-induced degradation of p-IκBα in DKO cells. B Immunoblotting of HA (CAND1 or CAND2), IκBα, and GAPDH at the indicated time points after the addition of TNFα to the indicated cells in the presence of cycloheximide (CHX). C Quantification of the relative p-IκBα levels in (B). Data are presented as mean ± SEM; n = 3 independent experiments. D, E Both CAND1 and CAND2 are required for proper degradation of IRP2 in HEK293 cells.

Journal: Nature communications

Article Title: Molecular mechanisms of CAND2 in regulating SCF ubiquitin ligases.

doi: 10.1038/s41467-025-57065-5

Figure Lengend Snippet: Fig. 1 | CAND2 promotes SCF-mediated protein degradation. A CAND2 binds to unneddylated CUL1 in human cell lysates. Whole-cell lysates (WCL) from HEK293 cells of the indicated genotypes were immunoprecipitated (IP) with an anti-HA antibody. Cells were treated with 1 μM MLN4924 for 1 h before harvesting. Samples were immunoblotted with anti-CUL1, anti-HA, and anti-GAPDH antibodies. DKO: CAND1 and CAND2 double knockout cells; DKOCAND1-HA: DKO cells expressing transgenic CAND1HA; DKOCAND2-HA: DKO cells expressing transgenic CAND2HA. Representative results from three independent experiments are shown. B, C CAND2 rescues the TNFα-induced degradation of p-IκBα in DKO cells. B Immunoblotting of HA (CAND1 or CAND2), IκBα, and GAPDH at the indicated time points after the addition of TNFα to the indicated cells in the presence of cycloheximide (CHX). C Quantification of the relative p-IκBα levels in (B). Data are presented as mean ± SEM; n = 3 independent experiments. D, E Both CAND1 and CAND2 are required for proper degradation of IRP2 in HEK293 cells.

Article Snippet: 4 s1 0.29 s-1 8.5 x 10-7 s-1 CUL1 CAND1S R CUL1 CAND2S R K M = 23 1 nM K M = 64 8 nM SR CAND1 CUL1 SR CAND2 CUL1 Nature Communications | (2025) 16:1998 11 Biotechnology # sc-47724, 1:5000), anti-CAND1 (Santa Cruz Biotechnology # 10672, 1:2000; Bethyl Laboratories # A302-901A, 1:1000), anti-CAND2 (Bethyl Laboratories # A304-046A, 1:1000), anti-CUL1 (Invitrogen # 32-2400, 1:500; Bethyl Laboratories # A303-373A, 1:500), anti-CUL2 (Thermo Fisher Scientific # 51-1800, 1:1000), antiCUL3 (Cell Signaling # 2759, 1:500), anti-CUL4A (Cell Signaling # 2699, 1:1000), anti-BCL10 (Santa Cruz Biotechnology # sc-5273, 1:200), antiSKP1 (Cell Signaling # 12248, 1:1000), anti-SKP2 (Cell Signaling # 2652, 1:1000), anti-HA (Cell Signaling # 3724, 1:1000), anti-FLAG (Sigma‒ Aldrich # F1804, 1:5000), anti-StrepII (Abcam # ab76949, 1:500), and anti-IκBα (Abcam#ab32518, 1:1000).

Techniques: Immunoprecipitation, Double Knockout, Expressing, Transgenic Assay, Western Blot