rabbit cc2d1b (Santa Cruz Biotechnology)
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Structured Review
Santa Cruz Biotechnology
rabbit cc2d1b
Rabbit Cc2d1b, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 88/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/rabbit+cc2d1b/CC2D1B+Antibody/pmc06286407-414-5-8
Average 88 stars, based on 1 article reviews
Rabbit Cc2d1b, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 88/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/rabbit+cc2d1b/CC2D1B+Antibody/pmc06286407-414-5-8
Average 88 stars, based on 1 article reviews
rabbit cc2d1b - by Bioz Stars,
2026-09
88/100 stars
Images
1) Product Images from "CC2D1B Coordinates ESCRT-III Activity during the Mitotic Reformation of the Nuclear Envelope"
Article Title: CC2D1B Coordinates ESCRT-III Activity during the Mitotic Reformation of the Nuclear Envelope
Journal: Developmental Cell
doi: 10.1016/j.devcel.2018.11.012
Figure Legend Snippet: HCT116 cells stably co-expressing GFP-NLS and H2B-mCherry were transfected with control or CC2D1B small interfering RNA (siRNA) and imaged every 5 min. The movie corresponding to siCC2D1B cells starts at 13 s.
Techniques Used:
Figure Legend Snippet: HCT116 δCC2D1B /GFP-CC2D1B cells were transfected with control or CHMP7 siRNA and imaged every 15 s. The movie corresponding to siCHMP7 cells starts at 6 s.
Techniques Used:
Figure Legend Snippet: HCT116 δ CC2D1B /GFP-CC2D1B cells stably expressing mCherry-Lap2β were imaged every 15 s.
Techniques Used:
Figure Legend Snippet: HCT116 δ CC2D1B /GFP-CC2D1B cells stably expressing mCherry-tubulin were imaged every 15 s.
Techniques Used:
Figure Legend Snippet: HCT116 δCC2D1B /GFP-CC2D1B Wt or HCT116 δCC2D1B /GFP-CC2D1B C2M cells were imaged every 15 s. The movie corresponding to GFP-CC2D1B C2M cells starts at 5 s.
Techniques Used:
Figure Legend Snippet: HCT116 cells stably expressing GFP-CHMP7 were transfected with control or CC2D1B siRNAs and imaged every 15 s. The movie corresponding to siCC2D1B cells starts at 10 s.
Techniques Used:
Figure Legend Snippet: HCT116 cells stably co-expressing CHMP4B-L-GFP and H2B-mCherry were transfected with control or CC2D1B siRNAs and imaged every 15 s. The movie corresponding to siCC2D1B cells starts at 8 s.
Techniques Used:
Figure Legend Snippet: HCT116 cells stably co-expressing CHMP2A-L-GFP and H2B-mCherry were transfected with control or CC2D1B siRNAs and imaged every 15 s. The movie corresponding to siCC2D1B cells starts at 10 s.
Techniques Used:
Figure Legend Snippet: HCT116 cells stably co-expressing GFP-NLS and H2B-mCherry were transfected with control of CC2D1B siRNAs and imaged every 1 min. The movie corresponding to siCC2D1B cells starts at 5 s.
Techniques Used:
Figure Legend Snippet: HCT116 cells stably co-expressing CHMP2A-L-mCherry and GFP-CC2D1B Wt or C2M were imaged every 15 s. The movie corresponding to GFP-CC2D1B C2M cells starts at 5 s.
Techniques Used:
Figure Legend Snippet: HeLa cells stably co-expressing YFP-Sec61β and H2B-mCherry were transfected with control or CC2D1B siRNAs and imaged every 15 s. The movie corresponding to siCC2D1B cells starts at 4 s.
Techniques Used:
Figure Legend Snippet: HCT116 cells stably co-expressing GFP-Spastin M87 and H2B-mCherry were transfected with control, CC2D1B, or CHMP7 siRNAs and imaged every 15 s. The movie corresponding to siCC2D1B cells starts at 8 s, and the movie corresponding to siCHMP7 cells starts at 17 s.
Techniques Used:
Figure Legend Snippet: HCT116 cells stably co-expressing CHMP2A-L-GFP and mCherry-tubulin were transfected with control, Spastin, or CC2D1B siRNAs and imaged every 15 s. The movie corresponding to siSpastin cells starts at 14 s, and the movie corresponding to siCC2D1B cells starts at 25 s.
Techniques Used:
Figure Legend Snippet: HCT116 cells stably co-expressing CHMP2A-L-GFP and mCherry-tubulin were transfected with control or CC2D1B siRNAs and Z-stacks with a separation of 0.1 μm between planes were acquired. The cells were incubated with Hoechst 33258 30 min before acquisition to stain DNA. The signal corresponding to CHMP2A-L-GFP is not shown. The signal corresponding to DNA is shown in green. The movie corresponding to siCC2D1B cells starts at 4 s.
Techniques Used:
Olmos et al., 2016 ), CHMP7 silencing was used to validate our experiments. (D) Representation of the nucleo-cytoplasmic GFP-NLS ratio over time. Mean ± SEM; siControl n = 29; siCC2D1B n = 35, p = 0.0003; siCHMP7 n = 22, p = 0.0402. Significance compared with the control was calculated at 90 min using a two-tailed unpaired t test. (E) Representative WB of total cell lysates corresponding to siCC2D1B or siCHMP7 treated cells. (F) Representative frames corresponding to time-lapse images of siControl (top panels) or siCC2D1B (bottom panels) treated cells. Scale bars, 10 μm. (G and H) Analysis of nuclear morphology in siRNA transfected HCT116 cells. (G) Percentage of cells showing an aberrant nuclear morphology. Mean ± SEM; siControl n = 2371; siCC2D1B n = 2040, p = 0.0102; siCHMP7 n = 1887, p = 0.0024. Significance of NE invaginations compared to the control was calculated using a two-tailed unpaired t test. (H) Representative images corresponding to the quantifications shown in (G). Scale bars, 10 μm. See also Figure Legend Snippet: CC2D1B Is Required to Maintain NE Integrity and Functionality (A) Schematic representation of CC2D1B and CHMP7. (B) GST pull-down experiments of 293T cells transiently co-expressing GST-CC2D1B together with HA-tagged CHMP2A, CHMP3, CHMP4B, or CHMP7. (C) GST pull-down experiments of 293T cells transiently co-expressing GST-CC2D1B together with HA-tagged CHMP7 full-length, CHMP7 Nt (residues 1–238), or CHMP7 Ct (residues 238–453). (D–F) Time-lapse analysis of HCT116 cells stably co-expressing GFP-NLS and H2B-mCherry and transfected with control, CHMP7 or CC2D1B siRNAs. As the recovery of nuclear integrity after cell division was previously described to be dependent on CHMP7 expression (
Techniques Used: Expressing, Stable Transfection, Transfection, Control, Two Tailed Test
Figure Legend Snippet: CC2D1B Is Recruited to the Reforming NE in a CHMP7-Dependent Way (A–C) Characterization of HCT116 δCC2D1B cells. (A) Total cell lysates corresponding to HCT116 Control, HCT116 δCC2D1B , HCT116 δCC2D1B /GFP and HCT116 δCC2D1B /GFP-CC2D1B were analyzed by WB using an anti-CC2D1B antibody. (B) Analysis of nuclear morphology in HCT116 δCC2D1B cells. Mean ± SEM; HCT116 Control n = 628; HCT116 δCC2D1B n = 608, p = 0.0162; HCT116 δCC2D1B /GFP n = 593, p = 0.0553; HCT116 δCC2D1B /GFP-CC2D1B n = 603, p = 0.9058. Significance of NE invaginations compared to the control was calculated using a two-tailed unpaired t test. (C) Representative images corresponding to the quantifications shown in (B). Scale bars, 10 μm. (D–F) Time-lapse images of HCT116 δCC2D1B /GFP-CC2D1B cells transfected with control (D) or CHMP7 siRNAs (E). Arrowheads indicate examples of GFP-CC2D1B perinuclear accumulation. Scale bars, 10 μm. (F) Representative WB corresponding to the cells shown in (D) and (E). (G) Time-lapse images of HCT116 δCC2D1B /GFP-CC2D1B cells stably co-expressing a mCherry-Lap2B fusion. Scale bar, 10 μm. (H) Time-lapse images of HCT116 δCC2D1B /GFP-CC2D1B cells stably co-expressing a mCherry-tubulin fusion. Scale bar, 10 μm. See also , , and .
Techniques Used: Control, Two Tailed Test, Transfection, Stable Transfection, Expressing
Figure S2 , , and . " title="... Is Required for Proper Localization and Functionality of CC2D1B (A) Schematic representation of Lgd. The arrow marks ..." property="contentUrl" width="100%" height="100%"/>
Figure Legend Snippet: A Functional C2 Domain Is Required for Proper Localization and Functionality of CC2D1B (A) Schematic representation of Lgd. The arrow marks the first residue of the solved structure. (B) Ribbon representation of Lgd C-terminal residues 575–816. The crystal structure was solved from a selenomethionine substituted crystal using the single wavelength anomalous dispersion method and the model refined to a resolution of 2.4 Å included the predicted C2 domain and the preceding region that lacks obvious sequence homology with known functional domains (residues 550–816). The helical domain composed of the N- and C-terminal region is shown in yellow and the C2 domain in orange. Conserved basic residues implicated in PIns(4,5)P2 interaction are shown as sticks. The N-terminal 24 residues connecting to the DM14 4 domain and present in the crystallized construct are flexible and disordered in the structure. Arrows indicate the site of insertion of the helical domain into the C2 domain. (C) Topology diagram of Lgd (residues 575–816) highlighting the insertion of the C2 domain into the helical domain. (D) Homology model of CC2D1B based on Lgd structure and docking of PIns(1,4,5)P 3 . The basic residues involved in PIns binding are shown as sticks. (E) Total cell lysates corresponding to HCT116 Control, HCT116 δCC2D1B , HCT116 δCC2D1B /GFP-CC2D1B Wt, and HCT116 δCC2D1B /GFP-CC2D1B C2M were analyzed by WB using an anti-CC2D1B antibody. (F) Time-lapse analysis of HCT116 δCC2D1B /GFP-CC2D1B Wt (top panels) or C2M (bottom panels) cells. Scale bars, 10 μm. (G) Nuclear inset of an HCT116 cell stably expressing CHMP4B-L-GFP and stained with anti-GFP, -PIP2 and -Lamin B1 antibodies. (H and I) Functional analysis of HCT116 δCC2D1B /GFP-CC2D1B C2M cells. (H) Representation of the nucleo-cytoplasmic ratio of mCherry-NLS over time in HCT116 δCC2D1B cells stably co-expressing GFP-CC2D1B Wt or C2M. Cells were incubated with Hoechst 33258 30 min before imaging to stain DNA. Mean ± SEM; HCT116 Control n = 14; HCT116 δCC2D1B n = 25, p < 0.0001; HCT116 δCC2D1B /GFP-CC2D1B Wt n = 18, p = 0.0597; HCT116 δCC2D1B /GFP-CC2D1B C2M n = 20, p < 0.0001. Significance compared to the control was calculated at 90 min using a two-tailed unpaired t test. (I) Analysis of nuclear morphology of HCT116 δCC2D1B cells stably expressing GFP-CC2D1B Wt or C2M. Mean ± SEM; HCT116 Control n = 1,055; HCT116 δCC2D1B n = 911, p = 0.0025; HCT116 δCC2D1B /GFP-CC2D1B Wt n = 896, p = 0.0651; HCT116 δCC2D1B /GFP-CC2D1B C2M n = 976, p = 0.0030. Significance of NE invaginations compared to the control was calculated using a two-tailed unpaired t test. See also
Techniques Used: Functional Assay, Residue, Dispersion, Sequencing, Construct, Binding Assay, Control, Stable Transfection, Expressing, Staining, Incubation, Imaging, Two Tailed Test
Figure 4 D. T0 was set at the beginning of furrow ingression (FI). GFP-CC2D1B n = 29; Ti p = 0.0183; Tf p < 0.0001. Significance compared to the control was calculated using a two-tailed unpaired t test. (I) Analysis of GFP-CC2D1B C2M expression on ESCRT-III recruitment time. HCT116 cells co-expressing CHMP2A-L-mCherry and GFP-CC2D1B Wt or C2M were analyzed by time-lapse microscopy and the first (Ti) and the last (Tf) frames showing CHMP2A-L-mCherry accumulation in the perinuclear area were scored. Whiskers mark 5–95 percentiles. T0 was set at the beginning of furrow ingression (FI). GFP-CC2D1B Wt n = 10; GFP-CC2D1B C2M n = 13; Ti p = 0.0803; Tf p = 0.0638. Significance compared to the control was calculated using a two-tailed unpaired t test. See also Figure Legend Snippet: CC2D1B Organizes the Timely Recruitment of ESCRT-III Proteins to the Reforming NE (A–C) Time-lapse analysis of HCT116 cells stably expressing GFP-CHMP7 (A), or stably co-expressing CHMP4B-L-GFP (B) or CHMP2A-L-GFP (C) along with H2B-mCherry. Cells were transfected with control or CC2D1B siRNAs. Scale bars, 10 μm in (A), (B), and (C). (D) Quantification of GFP-CHMP7, CHMP4B-L-GFP, and CHMP2A-L-GFP recruitment to the reforming NE in HCT116 cells transfected with control (top panel) or CC2D1B (bottom panel) siRNAs. T0 was set at the beginning of furrow ingression (FI). Curves indicate mean ± SEM. GFP-CHMP7: siControl n = 7; siCC2D1B n = 7. CHMP4B-L-GFP: siControl n = 11; siCC2D1B n = 13. CHMP2A-L-GFP: siControl n = 15; siCC2D1B n = 15. (E and F) Recruitment of endogenous CHMP2A to the reforming NE in HCT116 fixed cells. (E) Representative images of HCT116 cells transfected with control or CC2D1B siRNAs and stained with anti-CHMP2A and anti-tubulin antibodies. (F) Quantification of the midzone distance in CHMP2A-positive cells. Mean ± SEM; siControl n = 29; siCC2D1B n = 32, p = 0.0004. Scale bars, 10 μm in (E). (G) Sucrose gradient analysis of MBP-CHMP4B ΔC-Alix polymerization in the presence of CC2D1B (residues 317–558). Upper panel, CC2D1B (residues 317–558) floats in the top fractions of the gradient consistent with being monomeric; second panel, MBP-CHMP4B ΔC-Alix is found in the top fractions (monomers) and in the bottom (polymers) fraction; third panel, TEV cleavage of monomeric MBP-CHMP4B ΔC-Alix induces polymerization as indicated by the band in the bottom fraction; lower panel, TEV cleavage of monomeric MBP-CHMP4B ΔC-Alix in the presence of CC2D1B (residues 317–558) retains CHMP4B ΔC-Alix in the monomer fractions. (H) Comparison of CC2D1B and CHMP4B recruitment times. HCT116 cells expressing GFP-CC2D1B or CHMP4B-L-GFP were analyzed by time-lapse microscopy and the first (Ti) and the last (Tf) frames showing GFP-CC2D1B or CHMP4B-L-GFP accumulation in the perinuclear area were scored. Whiskers mark 5–95 percentiles. The movies used to quantify CHMP4B-L-GFP Ti and Tf were the same used for the recruitment quantification shown in
Techniques Used: Stable Transfection, Expressing, Transfection, Control, Staining, Comparison, Time-lapse Microscopy, Two Tailed Test
Figure S5 and . " title="CC2D1B Coordinates NM Deposition with ESCRT-III Recruitment to the ..." property="contentUrl" width="100%" height="100%"/>
Figure Legend Snippet: CC2D1B Coordinates NM Deposition with ESCRT-III Recruitment to the Resealing NE (A) Time-lapse analysis of HeLa cells stably co-expressing YFP-Sec61β and H2B-mCherry. Cells were transfected with control or CC2D1B siRNAs. Scale bars, 10 μm. (B) Time-lapse analysis of HeLa cells stably co-expressing CHMP4B-L-GFP and H2B-mCherry. Cells were transfected with control or CC2D1B siRNAs. Scale bars, 10 μm. (C) Quantification of YFP-Sec61β (Top panel. Lines: mean ± SEM) or CHMP4B-L-GFP (Bottom panel. Curves: mean ± SEM) recruitment to the NE in HeLa cells transfected with control or CC2D1B siRNAs. T0 was set at the beginning of furrow ingression (FI). YFP-Sec61β recruitment: siControl n = 15; siCC2D1B n = 10. CHMP4B-L-GFP recruitment: siControl n = 11; siCC2D1B n = 14. (D) Super resolution time-lapse analysis of HCT116 cells stably co-expressing CHMP4B-L-GFP and mCherry-Emerin and transfected with control or CC2D1B siRNAs. The frames corresponding to the beginning of CHMP4B recruitment (CHMP4B Ti) to the reforming NE are shown. (E) Quantification of the percentage of NE reformation at the beginning of CHMP4B recruitment (CHMP4B Ti). Bars indicate median. siControl n = 9; siCC2D1B n = 6, p = 0.0027. Significance compared to the control was calculated using a two-tailed unpaired t test. See also
Techniques Used: Stable Transfection, Expressing, Transfection, Control, Two Tailed Test
Figure 6 B. Cells were transfected with control or CC2D1B siRNAs. Bar indicates median. p = 0.0008. Significance compared to the control was calculated using a two-tailed unpaired t test. (E) Time-lapse analysis of HCT116 cells stably co-expressing mCherry-tubulin with CHMP2A-L-GFP and transfected with control, Spastin, or CC2D1B siRNAs. Scale bars, 10 μm. (F–I) Quantification of nuclear or cytoplasmic tubulin fluorescence over time in HCT116 cells stably co-expressing mCherry-tubulin with CHMP2A-L-GFP and transfected with Spastin (F and G) or CC2D1B (H and I) siRNAs. T0 was set at the beginning of furrow ingression. siControl n = 10; siCC2D1B n = 8; siSpastin n = 9. (F and H) Nuclear tubulin fluorescence (lines indicate mean ± SEM) siSpastin p = 0.0158; siCC2D1B p = 0.0294. CHMP2A-L-GFP recruitment to the NE (curve indicates mean ± SEM) in control cells. (G and I) Cytoplasmic tubulin fluorescence (lines indicate mean ± SEM) siSpastin p = 0.0099; siCC2D1B p = 0.0007. Significance compared to the control was calculated at 600 s using a two-tailed unpaired t test. See also Figure Legend Snippet: CC2D1B Silencing Impairs Spastin Activity (A) Time-lapse analysis of HCT116 cells stably co-expressing GFP-Spastin M87 and H2B-mCherry. Cells were transfected with control, CC2D1B, or CHMP7 siRNAs. Scale bars, 10 μm. (B) GFP-Spastin M87 fluorescence recruitment to the reforming NE in HCT116 cells transfected with control or CC2D1B siRNAs. T0 was set at the beginning of furrow ingression (FI). Curves indicate mean ± SEM. siControl n = 18; siCC2D1B n = 30. (C) Representative WB corresponding to the cells shown in (A). (D) Area under the curves corresponding to the recruitment of GFP-Spastin M87 fluorescence to the reforming NE shown in
Techniques Used: Activity Assay, Stable Transfection, Expressing, Transfection, Control, Fluorescence, Two Tailed Test
Figure S7 and . " title="... cell stably co-expressing mCherry-tubulin with CHMP2A-L-GFP, transfected with CC2D1B siRNA, and incubated with Hoechst 33258 30 min ..." property="contentUrl" width="100%" height="100%"/>
Figure Legend Snippet: Persistent Transnuclear Microtubules Originate Deep Nuclear Invaginations in Interphase (A) z stack series corresponding to an HCT116 cell stably co-expressing mCherry-tubulin with CHMP2A-L-GFP, transfected with CC2D1B siRNA, and incubated with Hoechst 33258 30 min before imaging to stain DNA. The signal corresponding to Hoechst 33258 is shown in green. The signal corresponding to CHMP2A-L-GFP is not shown. This cell was selected as an example of a nucleus showing transnuclear microtubule tunnels. Scale bar 10 μm. (B) Frequency of transnuclear microtubule tunnels in HCT116 cells stably co-expressing mCherry-tubulin with CHMP2A-L-GFP and transfected with control, Spastin, or CC2D1B siRNAs. Mean ± SEM; siControl n = 1,083; siCC2D1B n = 850, p = 0.0145; siSpastin n = 1,075, p = 0.0007. Significance compared to the control was calculated using a two-tailed unpaired t test. (C and D) 3D rendering of the cell shown in (A). (C) Front view corresponding to Hoechst 33258 channel. (D) Top view corresponding to Hoechst 33258 and mCherry-tubulin. 80% transparency was applied to Hoechst 33258 channel to facilitate the observation of transnuclear microtubule channels. Scale bars, 5 μm. (E) Proposed model. Left panel: (1) CC2D1B binds to the monomeric cytoplasmic form of CHMP4B, hindering its association with CHMP7 and therefore preventing its premature recruitment to the NE. (2) By means of its ability to interact with CHMP7 and CHMP4B through its N-terminal DM14 domains and with membrane phospholipids through its C-terminal C2 domain, CC2D1B could function as a scaffold to position CHMP4B monomers close to the NM. Then, CC2D1B could mediate the organization of a transient CHMP7-CC2D1B-CHMP4B complex at the sealing gaps of the NE, facilitating the delivery of CHMP4B monomers to the growing ESCRT-III filament. (3) Spastin is recruited by the ESCRT-III to catalyze the severing of spindle microtubules, which facilitates the final sealing of the NE. Right panel: (1) in CC2D1B-silenced cells, CHMP4B monomers are free to associate to CHMP7 without restrictions, (2) triggering the premature polymerization of CHMP4B monomers on the NE. (3) These prematurely formed ESCRT filaments cannot be competent for Spastin recruitment and membrane constriction, which results in an impaired ability of the cells to sever spindle microtubules, leading to deleterious effects in nuclear integrity and functionality. See also
Techniques Used: Stable Transfection, Expressing, Transfection, Incubation, Imaging, Staining, Control, Two Tailed Test, Membrane
Figure Legend Snippet:
Techniques Used: Virus, Recombinant, Protease Inhibitor, Software
