mouse monoclonal anti p32 antibody  (Hycult Biotech)


Bioz Verified Symbol Hycult Biotech is a verified supplier
Bioz Manufacturer Symbol Hycult Biotech manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 90

    Structured Review

    Hycult Biotech mouse monoclonal anti p32 antibody
    Mouse Monoclonal Anti P32 Antibody, supplied by Hycult Biotech, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/mouse monoclonal anti p32 antibody/product/Hycult Biotech
    Average 90 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    mouse monoclonal anti p32 antibody - by Bioz Stars, 2024-05
    90/100 stars

    Images

    mouse monoclonal anti p32 antibody  (Hycult Biotech)


    Bioz Verified Symbol Hycult Biotech is a verified supplier
    Bioz Manufacturer Symbol Hycult Biotech manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 90

    Structured Review

    Hycult Biotech mouse monoclonal anti p32 antibody
    Mouse Monoclonal Anti P32 Antibody, supplied by Hycult Biotech, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/mouse monoclonal anti p32 antibody/product/Hycult Biotech
    Average 90 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    mouse monoclonal anti p32 antibody - by Bioz Stars, 2024-05
    90/100 stars

    Images

    mouse monoclonal anti p32 antibody  (Hycult Biotech)


    Bioz Verified Symbol Hycult Biotech is a verified supplier
    Bioz Manufacturer Symbol Hycult Biotech manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 90

    Structured Review

    Hycult Biotech mouse monoclonal anti p32 antibody
    <t>p32</t> deletion impairs the nuclear egress of PCV2 capsids. (A) Examination of p32 expression in PK-15 cells with a CRISPR/Cas9 system targeting the p32 locus. Three single-cell clones [p32(22), p32(155), and p32(173)] were derived from cells infected with lentiviral pseudotypes expressing gRNAs 22, 155, and 173, respectively. (B, C) Wild-type PK-15, 22PKp32−/−, 155PKp32−/−, and 173PKp32+/+ cells were infected with PCV2, supernatant and cells were collected at indicated times, and the titers of PCV2 in extracellular (B) and intracellular (C) were detected by 50% tissue culture infective dose (TCID50), respectively. *, P < 0.05. Comparison were made at the same time points between values in p32 knockout (KO) cells and wild-type PK-15 cells infected by PCV2. (D to E) Depletion of p32 blocks the nuclear egress of PCV2 capsids. Wild-type PK-15, 22PKp32−/−, 155PKp32−/−, and 173PKp32+/+ cells were infected with PCV2 for 24, 36. and 48 h, the proportion of viral DNA copies in cytoplasm and nucleus was measured by quantitative PCR (qPCR) (D), Cap levels were detected by Western blotting, and the relative Cap levels were calculated using ImageJ. (E) The proportion of Cap (nucleus/cytoplasm) is shown (E). *, P < 0.05; **, P < 0.01. Comparisons were made between values in p32 KO cells and wild-type PK-15 cells infected by PCV2 at the same time points; ##, P < 0.01 (compared with the percentage in cytoplasm). (F to G) 22PKp32−/− and 173PKp32+/+ cells were transfected with plasmids to express wild-type p32 (WT) or control vector pCI-neo (Vec), respectively, and then the cells were infected with PCV2; after 36 h, the proportions of Cap proteins (nucleus/cytoplasm) (F) and PCV2 DNA in nucleus or cytoplasm (G) were measured. *, P < 0.05 (compared with 173PKp32+/+ cells transfected with Vec); ##, P < 0.01 (compared with percentage in cytoplasm).
    Mouse Monoclonal Anti P32 Antibody, supplied by Hycult Biotech, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/mouse monoclonal anti p32 antibody/product/Hycult Biotech
    Average 90 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    mouse monoclonal anti p32 antibody - by Bioz Stars, 2024-05
    90/100 stars

    Images

    1) Product Images from "Cellular p32 Is a Critical Regulator of Porcine Circovirus Type 2 Nuclear Egress"

    Article Title: Cellular p32 Is a Critical Regulator of Porcine Circovirus Type 2 Nuclear Egress

    Journal: Journal of Virology

    doi: 10.1128/JVI.00979-19

    p32 deletion impairs the nuclear egress of PCV2 capsids. (A) Examination of p32 expression in PK-15 cells with a CRISPR/Cas9 system targeting the p32 locus. Three single-cell clones [p32(22), p32(155), and p32(173)] were derived from cells infected with lentiviral pseudotypes expressing gRNAs 22, 155, and 173, respectively. (B, C) Wild-type PK-15, 22PKp32−/−, 155PKp32−/−, and 173PKp32+/+ cells were infected with PCV2, supernatant and cells were collected at indicated times, and the titers of PCV2 in extracellular (B) and intracellular (C) were detected by 50% tissue culture infective dose (TCID50), respectively. *, P < 0.05. Comparison were made at the same time points between values in p32 knockout (KO) cells and wild-type PK-15 cells infected by PCV2. (D to E) Depletion of p32 blocks the nuclear egress of PCV2 capsids. Wild-type PK-15, 22PKp32−/−, 155PKp32−/−, and 173PKp32+/+ cells were infected with PCV2 for 24, 36. and 48 h, the proportion of viral DNA copies in cytoplasm and nucleus was measured by quantitative PCR (qPCR) (D), Cap levels were detected by Western blotting, and the relative Cap levels were calculated using ImageJ. (E) The proportion of Cap (nucleus/cytoplasm) is shown (E). *, P < 0.05; **, P < 0.01. Comparisons were made between values in p32 KO cells and wild-type PK-15 cells infected by PCV2 at the same time points; ##, P < 0.01 (compared with the percentage in cytoplasm). (F to G) 22PKp32−/− and 173PKp32+/+ cells were transfected with plasmids to express wild-type p32 (WT) or control vector pCI-neo (Vec), respectively, and then the cells were infected with PCV2; after 36 h, the proportions of Cap proteins (nucleus/cytoplasm) (F) and PCV2 DNA in nucleus or cytoplasm (G) were measured. *, P < 0.05 (compared with 173PKp32+/+ cells transfected with Vec); ##, P < 0.01 (compared with percentage in cytoplasm).
    Figure Legend Snippet: p32 deletion impairs the nuclear egress of PCV2 capsids. (A) Examination of p32 expression in PK-15 cells with a CRISPR/Cas9 system targeting the p32 locus. Three single-cell clones [p32(22), p32(155), and p32(173)] were derived from cells infected with lentiviral pseudotypes expressing gRNAs 22, 155, and 173, respectively. (B, C) Wild-type PK-15, 22PKp32−/−, 155PKp32−/−, and 173PKp32+/+ cells were infected with PCV2, supernatant and cells were collected at indicated times, and the titers of PCV2 in extracellular (B) and intracellular (C) were detected by 50% tissue culture infective dose (TCID50), respectively. *, P < 0.05. Comparison were made at the same time points between values in p32 knockout (KO) cells and wild-type PK-15 cells infected by PCV2. (D to E) Depletion of p32 blocks the nuclear egress of PCV2 capsids. Wild-type PK-15, 22PKp32−/−, 155PKp32−/−, and 173PKp32+/+ cells were infected with PCV2 for 24, 36. and 48 h, the proportion of viral DNA copies in cytoplasm and nucleus was measured by quantitative PCR (qPCR) (D), Cap levels were detected by Western blotting, and the relative Cap levels were calculated using ImageJ. (E) The proportion of Cap (nucleus/cytoplasm) is shown (E). *, P < 0.05; **, P < 0.01. Comparisons were made between values in p32 KO cells and wild-type PK-15 cells infected by PCV2 at the same time points; ##, P < 0.01 (compared with the percentage in cytoplasm). (F to G) 22PKp32−/− and 173PKp32+/+ cells were transfected with plasmids to express wild-type p32 (WT) or control vector pCI-neo (Vec), respectively, and then the cells were infected with PCV2; after 36 h, the proportions of Cap proteins (nucleus/cytoplasm) (F) and PCV2 DNA in nucleus or cytoplasm (G) were measured. *, P < 0.05 (compared with 173PKp32+/+ cells transfected with Vec); ##, P < 0.01 (compared with percentage in cytoplasm).

    Techniques Used: Expressing, CRISPR, Clone Assay, Derivative Assay, Infection, Knock-Out, Real-time Polymerase Chain Reaction, Western Blot, Transfection, Plasmid Preparation

    PCV2 infection induces the phosphorylation and redistribution of lamin A/C mediated by p32. (A, B) p32 knockout promoted the redistribution of lamin A/C and reduced the phosphorylation of lamin A/C. Wild-type PK-15, 173PKp32+/+, and 22PKp32−/− cells were infected with PCV2, and the redistribution of lamin A/C was determined by laser scanning confocal microscopy (A); the phosphorylation levels of lamin A/C were measured by Western blotting (B).
    Figure Legend Snippet: PCV2 infection induces the phosphorylation and redistribution of lamin A/C mediated by p32. (A, B) p32 knockout promoted the redistribution of lamin A/C and reduced the phosphorylation of lamin A/C. Wild-type PK-15, 173PKp32+/+, and 22PKp32−/− cells were infected with PCV2, and the redistribution of lamin A/C was determined by laser scanning confocal microscopy (A); the phosphorylation levels of lamin A/C were measured by Western blotting (B).

    Techniques Used: Infection, Knock-Out, Confocal Microscopy, Western Blot

    p32 mediated the interaction of lamin A/C, lamin B receptor (LBR), and Cap. (A) PK-15 cells were transfected with pEGFP-Cap and pCI-p32-Flag vectors or with pCI-p32-Flag and pEGFP-N1 vectors; the cells were fixed and subjected to laser scanning confocal microscopy. Images represent the subcellular locations of green fluorescent protein (GFP)-Cap and Flag-p32 proteins (left), and histograms represent the percentage of overlap of Flag-p32 proteins with GFP-Cap, performed using ImageJ software and based on ≥15 cells/sample (right). **, P < 0.01 (compared with pCI-p32-Flag and pEGFP-N1 vector cotransfected cells). (B, C) p32 mediates the interaction of lamin A/C, LBR, and Cap protein. Wild-type PK-15, 173PKp32+/+, and 22PKp32−/− cells were infected with PCV2, and immunoprecipitation was performed to detect the Cap interaction with lamin A/C, LBR, and p32 using anti-Cap antibodies (B) or anti-lamin A/C antibodies (C). (D to F) Direct interaction of p32 with PCV2 Cap or LBR. Bacterially purified GST-p32 or glutathione S-transferase (GST) alone was incubated with purified His-Cap, and proteins bound to glutathione Sepharose beads were analyzed by immunoblotting with the indicated antibodies (D); purified GST-LBR or GST alone was incubated with purified His-p32 (E) or His-Cap (F). Proteins bound to glutathione Sepharose beads were analyzed by immunoblotting with the indicated antibodies.
    Figure Legend Snippet: p32 mediated the interaction of lamin A/C, lamin B receptor (LBR), and Cap. (A) PK-15 cells were transfected with pEGFP-Cap and pCI-p32-Flag vectors or with pCI-p32-Flag and pEGFP-N1 vectors; the cells were fixed and subjected to laser scanning confocal microscopy. Images represent the subcellular locations of green fluorescent protein (GFP)-Cap and Flag-p32 proteins (left), and histograms represent the percentage of overlap of Flag-p32 proteins with GFP-Cap, performed using ImageJ software and based on ≥15 cells/sample (right). **, P < 0.01 (compared with pCI-p32-Flag and pEGFP-N1 vector cotransfected cells). (B, C) p32 mediates the interaction of lamin A/C, LBR, and Cap protein. Wild-type PK-15, 173PKp32+/+, and 22PKp32−/− cells were infected with PCV2, and immunoprecipitation was performed to detect the Cap interaction with lamin A/C, LBR, and p32 using anti-Cap antibodies (B) or anti-lamin A/C antibodies (C). (D to F) Direct interaction of p32 with PCV2 Cap or LBR. Bacterially purified GST-p32 or glutathione S-transferase (GST) alone was incubated with purified His-Cap, and proteins bound to glutathione Sepharose beads were analyzed by immunoblotting with the indicated antibodies (D); purified GST-LBR or GST alone was incubated with purified His-p32 (E) or His-Cap (F). Proteins bound to glutathione Sepharose beads were analyzed by immunoblotting with the indicated antibodies.

    Techniques Used: Transfection, Confocal Microscopy, Software, Plasmid Preparation, Infection, Immunoprecipitation, Purification, Incubation, Western Blot

    p32 recruits PKC-δ to the nuclear membrane to phosphorylate lamin A/C during PCV2 infection. (A) Direct interaction of PKC-δ with Cap or p32. Purified GST-PKC-δ or GST alone was incubated with purified His-Cap (left panel) or His-p32 (right panel), and proteins bound to glutathione Sepharose beads were analyzed by immunoblotting with the indicated antibodies. (B) Knockout of p32 reduced the phosphorylation levels of PKC-δ (T505). Wild type PK-15 and 22PKp32−/− cells were infected with PCV2, the cell lysates were subjected to immunoblotting using anti-PKC-δ (T505) and PKC-δ antibodies. (C, D) p32 deficiency impeded recruitment of p-PKC-δ to nuclear membrane during PCV2 infection. Wild-type PK-15, 173PKp32+/+, and 22PKp32−/− cells were infected with PCV2. The interaction of p-PKC-δ (T505) with Cap (C) or lamin A/C (D) and the distribution of these molecules were observed under laser scanning confocal microscopy.
    Figure Legend Snippet: p32 recruits PKC-δ to the nuclear membrane to phosphorylate lamin A/C during PCV2 infection. (A) Direct interaction of PKC-δ with Cap or p32. Purified GST-PKC-δ or GST alone was incubated with purified His-Cap (left panel) or His-p32 (right panel), and proteins bound to glutathione Sepharose beads were analyzed by immunoblotting with the indicated antibodies. (B) Knockout of p32 reduced the phosphorylation levels of PKC-δ (T505). Wild type PK-15 and 22PKp32−/− cells were infected with PCV2, the cell lysates were subjected to immunoblotting using anti-PKC-δ (T505) and PKC-δ antibodies. (C, D) p32 deficiency impeded recruitment of p-PKC-δ to nuclear membrane during PCV2 infection. Wild-type PK-15, 173PKp32+/+, and 22PKp32−/− cells were infected with PCV2. The interaction of p-PKC-δ (T505) with Cap (C) or lamin A/C (D) and the distribution of these molecules were observed under laser scanning confocal microscopy.

    Techniques Used: Infection, Purification, Incubation, Western Blot, Knock-Out, Confocal Microscopy

    Cap binds to p32 via its N-terminal 24RRR26. (A) Cap fragment (aa 1 to 41) interacted with p32. HEK293T cells were cotransfected with plasmids encoding full-length Cap or deletion mutants fused with a GFP tag, along with Flag-p32; cell lysates were immunoprecipitated with an anti-GFP antibody and immunoblotted using indicated antibodies. (B) The Cap N-terminal aa residues 21 to 41 interacted with p32. HEK293T cells were cotransfected with plasmids encoding the N-terminal aa residues 1 to 41, 1 to 20, or 21 to 41 fused with a GFP tag, along with Flag-p32; cell lysates were immunoprecipitated with anti-GFP antibody, followed by immunoblotting using indicated antibodies. (C) The Cap mutants were constructed with arginine residues at residues 24 to 26, 33, 35, and 37 replaced with alanine. (D) Mapping the crucial amino acids of Cap responsible for Cap/p32 interaction. HEK293T cells were cotransfected with Cap or Cap mutants 1, 2, and 3, along with Flag-p32 expression vectors, and the cell lysates were subjected to immunoprecipitation and immunoblotting using the indicated antibodies. (E) Confirmation by immunofluorescence assay of the crucial amino acids of PCV2 Cap that interact with p32. PK-15 cells were cotransfected with plasmids indicated in panel D and subjected to confocal microscopy observation at 36 h posttransfection. These results were confirmed in three independent experiments.
    Figure Legend Snippet: Cap binds to p32 via its N-terminal 24RRR26. (A) Cap fragment (aa 1 to 41) interacted with p32. HEK293T cells were cotransfected with plasmids encoding full-length Cap or deletion mutants fused with a GFP tag, along with Flag-p32; cell lysates were immunoprecipitated with an anti-GFP antibody and immunoblotted using indicated antibodies. (B) The Cap N-terminal aa residues 21 to 41 interacted with p32. HEK293T cells were cotransfected with plasmids encoding the N-terminal aa residues 1 to 41, 1 to 20, or 21 to 41 fused with a GFP tag, along with Flag-p32; cell lysates were immunoprecipitated with anti-GFP antibody, followed by immunoblotting using indicated antibodies. (C) The Cap mutants were constructed with arginine residues at residues 24 to 26, 33, 35, and 37 replaced with alanine. (D) Mapping the crucial amino acids of Cap responsible for Cap/p32 interaction. HEK293T cells were cotransfected with Cap or Cap mutants 1, 2, and 3, along with Flag-p32 expression vectors, and the cell lysates were subjected to immunoprecipitation and immunoblotting using the indicated antibodies. (E) Confirmation by immunofluorescence assay of the crucial amino acids of PCV2 Cap that interact with p32. PK-15 cells were cotransfected with plasmids indicated in panel D and subjected to confocal microscopy observation at 36 h posttransfection. These results were confirmed in three independent experiments.

    Techniques Used: Immunoprecipitation, Western Blot, Construct, Expressing, Immunofluorescence, Confocal Microscopy

    Model of PCV2 nuclear egress. PCV2 infection promotes the phosphorylation of PKC-δ via PLC-mediated signaling at the early infection phase, which is further amplified by JNK and ERK1/2 signaling at the late infection phase. In wild-type cells, Cap recruits p32 and phosphorylates PKC-δ (p-PKC-δ), and this leads to transport of p32 and p-PKC-δ into the nucleus. In the nucleus, p32, as an adaptor, further recruits p-PKC-δ and Cap to the nuclear membrane via binding with LBR to phosphorylate lamin A/C, which promotes the rearrangement of nuclear lamina and facilitates viral nuclear egress.
    Figure Legend Snippet: Model of PCV2 nuclear egress. PCV2 infection promotes the phosphorylation of PKC-δ via PLC-mediated signaling at the early infection phase, which is further amplified by JNK and ERK1/2 signaling at the late infection phase. In wild-type cells, Cap recruits p32 and phosphorylates PKC-δ (p-PKC-δ), and this leads to transport of p32 and p-PKC-δ into the nucleus. In the nucleus, p32, as an adaptor, further recruits p-PKC-δ and Cap to the nuclear membrane via binding with LBR to phosphorylate lamin A/C, which promotes the rearrangement of nuclear lamina and facilitates viral nuclear egress.

    Techniques Used: Infection, Amplification, Binding Assay

    mouse monoclonal anti p32 antibody  (Hycult Biotech)


    Bioz Verified Symbol Hycult Biotech is a verified supplier
    Bioz Manufacturer Symbol Hycult Biotech manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 90

    Structured Review

    Hycult Biotech mouse monoclonal anti p32 antibody
    <t>p32</t> deletion impairs the nuclear egress of PCV2 capsids. (A) Examination of p32 expression in PK-15 cells with a CRISPR/Cas9 system targeting the p32 locus. Three single-cell clones [p32(22), p32(155), and p32(173)] were derived from cells infected with lentiviral pseudotypes expressing gRNAs 22, 155, and 173, respectively. (B, C) Wild-type PK-15, 22PKp32−/−, 155PKp32−/−, and 173PKp32+/+ cells were infected with PCV2, supernatant and cells were collected at indicated times, and the titers of PCV2 in extracellular (B) and intracellular (C) were detected by 50% tissue culture infective dose (TCID50), respectively. *, P < 0.05. Comparison were made at the same time points between values in p32 knockout (KO) cells and wild-type PK-15 cells infected by PCV2. (D to E) Depletion of p32 blocks the nuclear egress of PCV2 capsids. Wild-type PK-15, 22PKp32−/−, 155PKp32−/−, and 173PKp32+/+ cells were infected with PCV2 for 24, 36. and 48 h, the proportion of viral DNA copies in cytoplasm and nucleus was measured by quantitative PCR (qPCR) (D), Cap levels were detected by Western blotting, and the relative Cap levels were calculated using ImageJ. (E) The proportion of Cap (nucleus/cytoplasm) is shown (E). *, P < 0.05; **, P < 0.01. Comparisons were made between values in p32 KO cells and wild-type PK-15 cells infected by PCV2 at the same time points; ##, P < 0.01 (compared with the percentage in cytoplasm). (F to G) 22PKp32−/− and 173PKp32+/+ cells were transfected with plasmids to express wild-type p32 (WT) or control vector pCI-neo (Vec), respectively, and then the cells were infected with PCV2; after 36 h, the proportions of Cap proteins (nucleus/cytoplasm) (F) and PCV2 DNA in nucleus or cytoplasm (G) were measured. *, P < 0.05 (compared with 173PKp32+/+ cells transfected with Vec); ##, P < 0.01 (compared with percentage in cytoplasm).
    Mouse Monoclonal Anti P32 Antibody, supplied by Hycult Biotech, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/mouse monoclonal anti p32 antibody/product/Hycult Biotech
    Average 90 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    mouse monoclonal anti p32 antibody - by Bioz Stars, 2024-05
    90/100 stars

    Images

    1) Product Images from "Cellular p32 Is a Critical Regulator of Porcine Circovirus Type 2 Nuclear Egress"

    Article Title: Cellular p32 Is a Critical Regulator of Porcine Circovirus Type 2 Nuclear Egress

    Journal: Journal of Virology

    doi: 10.1128/JVI.00979-19

    p32 deletion impairs the nuclear egress of PCV2 capsids. (A) Examination of p32 expression in PK-15 cells with a CRISPR/Cas9 system targeting the p32 locus. Three single-cell clones [p32(22), p32(155), and p32(173)] were derived from cells infected with lentiviral pseudotypes expressing gRNAs 22, 155, and 173, respectively. (B, C) Wild-type PK-15, 22PKp32−/−, 155PKp32−/−, and 173PKp32+/+ cells were infected with PCV2, supernatant and cells were collected at indicated times, and the titers of PCV2 in extracellular (B) and intracellular (C) were detected by 50% tissue culture infective dose (TCID50), respectively. *, P < 0.05. Comparison were made at the same time points between values in p32 knockout (KO) cells and wild-type PK-15 cells infected by PCV2. (D to E) Depletion of p32 blocks the nuclear egress of PCV2 capsids. Wild-type PK-15, 22PKp32−/−, 155PKp32−/−, and 173PKp32+/+ cells were infected with PCV2 for 24, 36. and 48 h, the proportion of viral DNA copies in cytoplasm and nucleus was measured by quantitative PCR (qPCR) (D), Cap levels were detected by Western blotting, and the relative Cap levels were calculated using ImageJ. (E) The proportion of Cap (nucleus/cytoplasm) is shown (E). *, P < 0.05; **, P < 0.01. Comparisons were made between values in p32 KO cells and wild-type PK-15 cells infected by PCV2 at the same time points; ##, P < 0.01 (compared with the percentage in cytoplasm). (F to G) 22PKp32−/− and 173PKp32+/+ cells were transfected with plasmids to express wild-type p32 (WT) or control vector pCI-neo (Vec), respectively, and then the cells were infected with PCV2; after 36 h, the proportions of Cap proteins (nucleus/cytoplasm) (F) and PCV2 DNA in nucleus or cytoplasm (G) were measured. *, P < 0.05 (compared with 173PKp32+/+ cells transfected with Vec); ##, P < 0.01 (compared with percentage in cytoplasm).
    Figure Legend Snippet: p32 deletion impairs the nuclear egress of PCV2 capsids. (A) Examination of p32 expression in PK-15 cells with a CRISPR/Cas9 system targeting the p32 locus. Three single-cell clones [p32(22), p32(155), and p32(173)] were derived from cells infected with lentiviral pseudotypes expressing gRNAs 22, 155, and 173, respectively. (B, C) Wild-type PK-15, 22PKp32−/−, 155PKp32−/−, and 173PKp32+/+ cells were infected with PCV2, supernatant and cells were collected at indicated times, and the titers of PCV2 in extracellular (B) and intracellular (C) were detected by 50% tissue culture infective dose (TCID50), respectively. *, P < 0.05. Comparison were made at the same time points between values in p32 knockout (KO) cells and wild-type PK-15 cells infected by PCV2. (D to E) Depletion of p32 blocks the nuclear egress of PCV2 capsids. Wild-type PK-15, 22PKp32−/−, 155PKp32−/−, and 173PKp32+/+ cells were infected with PCV2 for 24, 36. and 48 h, the proportion of viral DNA copies in cytoplasm and nucleus was measured by quantitative PCR (qPCR) (D), Cap levels were detected by Western blotting, and the relative Cap levels were calculated using ImageJ. (E) The proportion of Cap (nucleus/cytoplasm) is shown (E). *, P < 0.05; **, P < 0.01. Comparisons were made between values in p32 KO cells and wild-type PK-15 cells infected by PCV2 at the same time points; ##, P < 0.01 (compared with the percentage in cytoplasm). (F to G) 22PKp32−/− and 173PKp32+/+ cells were transfected with plasmids to express wild-type p32 (WT) or control vector pCI-neo (Vec), respectively, and then the cells were infected with PCV2; after 36 h, the proportions of Cap proteins (nucleus/cytoplasm) (F) and PCV2 DNA in nucleus or cytoplasm (G) were measured. *, P < 0.05 (compared with 173PKp32+/+ cells transfected with Vec); ##, P < 0.01 (compared with percentage in cytoplasm).

    Techniques Used: Expressing, CRISPR, Clone Assay, Derivative Assay, Infection, Knock-Out, Real-time Polymerase Chain Reaction, Western Blot, Transfection, Plasmid Preparation

    PCV2 infection induces the phosphorylation and redistribution of lamin A/C mediated by p32. (A, B) p32 knockout promoted the redistribution of lamin A/C and reduced the phosphorylation of lamin A/C. Wild-type PK-15, 173PKp32+/+, and 22PKp32−/− cells were infected with PCV2, and the redistribution of lamin A/C was determined by laser scanning confocal microscopy (A); the phosphorylation levels of lamin A/C were measured by Western blotting (B).
    Figure Legend Snippet: PCV2 infection induces the phosphorylation and redistribution of lamin A/C mediated by p32. (A, B) p32 knockout promoted the redistribution of lamin A/C and reduced the phosphorylation of lamin A/C. Wild-type PK-15, 173PKp32+/+, and 22PKp32−/− cells were infected with PCV2, and the redistribution of lamin A/C was determined by laser scanning confocal microscopy (A); the phosphorylation levels of lamin A/C were measured by Western blotting (B).

    Techniques Used: Infection, Knock-Out, Confocal Microscopy, Western Blot

    p32 mediated the interaction of lamin A/C, lamin B receptor (LBR), and Cap. (A) PK-15 cells were transfected with pEGFP-Cap and pCI-p32-Flag vectors or with pCI-p32-Flag and pEGFP-N1 vectors; the cells were fixed and subjected to laser scanning confocal microscopy. Images represent the subcellular locations of green fluorescent protein (GFP)-Cap and Flag-p32 proteins (left), and histograms represent the percentage of overlap of Flag-p32 proteins with GFP-Cap, performed using ImageJ software and based on ≥15 cells/sample (right). **, P < 0.01 (compared with pCI-p32-Flag and pEGFP-N1 vector cotransfected cells). (B, C) p32 mediates the interaction of lamin A/C, LBR, and Cap protein. Wild-type PK-15, 173PKp32+/+, and 22PKp32−/− cells were infected with PCV2, and immunoprecipitation was performed to detect the Cap interaction with lamin A/C, LBR, and p32 using anti-Cap antibodies (B) or anti-lamin A/C antibodies (C). (D to F) Direct interaction of p32 with PCV2 Cap or LBR. Bacterially purified GST-p32 or glutathione S-transferase (GST) alone was incubated with purified His-Cap, and proteins bound to glutathione Sepharose beads were analyzed by immunoblotting with the indicated antibodies (D); purified GST-LBR or GST alone was incubated with purified His-p32 (E) or His-Cap (F). Proteins bound to glutathione Sepharose beads were analyzed by immunoblotting with the indicated antibodies.
    Figure Legend Snippet: p32 mediated the interaction of lamin A/C, lamin B receptor (LBR), and Cap. (A) PK-15 cells were transfected with pEGFP-Cap and pCI-p32-Flag vectors or with pCI-p32-Flag and pEGFP-N1 vectors; the cells were fixed and subjected to laser scanning confocal microscopy. Images represent the subcellular locations of green fluorescent protein (GFP)-Cap and Flag-p32 proteins (left), and histograms represent the percentage of overlap of Flag-p32 proteins with GFP-Cap, performed using ImageJ software and based on ≥15 cells/sample (right). **, P < 0.01 (compared with pCI-p32-Flag and pEGFP-N1 vector cotransfected cells). (B, C) p32 mediates the interaction of lamin A/C, LBR, and Cap protein. Wild-type PK-15, 173PKp32+/+, and 22PKp32−/− cells were infected with PCV2, and immunoprecipitation was performed to detect the Cap interaction with lamin A/C, LBR, and p32 using anti-Cap antibodies (B) or anti-lamin A/C antibodies (C). (D to F) Direct interaction of p32 with PCV2 Cap or LBR. Bacterially purified GST-p32 or glutathione S-transferase (GST) alone was incubated with purified His-Cap, and proteins bound to glutathione Sepharose beads were analyzed by immunoblotting with the indicated antibodies (D); purified GST-LBR or GST alone was incubated with purified His-p32 (E) or His-Cap (F). Proteins bound to glutathione Sepharose beads were analyzed by immunoblotting with the indicated antibodies.

    Techniques Used: Transfection, Confocal Microscopy, Software, Plasmid Preparation, Infection, Immunoprecipitation, Purification, Incubation, Western Blot

    p32 recruits PKC-δ to the nuclear membrane to phosphorylate lamin A/C during PCV2 infection. (A) Direct interaction of PKC-δ with Cap or p32. Purified GST-PKC-δ or GST alone was incubated with purified His-Cap (left panel) or His-p32 (right panel), and proteins bound to glutathione Sepharose beads were analyzed by immunoblotting with the indicated antibodies. (B) Knockout of p32 reduced the phosphorylation levels of PKC-δ (T505). Wild type PK-15 and 22PKp32−/− cells were infected with PCV2, the cell lysates were subjected to immunoblotting using anti-PKC-δ (T505) and PKC-δ antibodies. (C, D) p32 deficiency impeded recruitment of p-PKC-δ to nuclear membrane during PCV2 infection. Wild-type PK-15, 173PKp32+/+, and 22PKp32−/− cells were infected with PCV2. The interaction of p-PKC-δ (T505) with Cap (C) or lamin A/C (D) and the distribution of these molecules were observed under laser scanning confocal microscopy.
    Figure Legend Snippet: p32 recruits PKC-δ to the nuclear membrane to phosphorylate lamin A/C during PCV2 infection. (A) Direct interaction of PKC-δ with Cap or p32. Purified GST-PKC-δ or GST alone was incubated with purified His-Cap (left panel) or His-p32 (right panel), and proteins bound to glutathione Sepharose beads were analyzed by immunoblotting with the indicated antibodies. (B) Knockout of p32 reduced the phosphorylation levels of PKC-δ (T505). Wild type PK-15 and 22PKp32−/− cells were infected with PCV2, the cell lysates were subjected to immunoblotting using anti-PKC-δ (T505) and PKC-δ antibodies. (C, D) p32 deficiency impeded recruitment of p-PKC-δ to nuclear membrane during PCV2 infection. Wild-type PK-15, 173PKp32+/+, and 22PKp32−/− cells were infected with PCV2. The interaction of p-PKC-δ (T505) with Cap (C) or lamin A/C (D) and the distribution of these molecules were observed under laser scanning confocal microscopy.

    Techniques Used: Infection, Purification, Incubation, Western Blot, Knock-Out, Confocal Microscopy

    Cap binds to p32 via its N-terminal 24RRR26. (A) Cap fragment (aa 1 to 41) interacted with p32. HEK293T cells were cotransfected with plasmids encoding full-length Cap or deletion mutants fused with a GFP tag, along with Flag-p32; cell lysates were immunoprecipitated with an anti-GFP antibody and immunoblotted using indicated antibodies. (B) The Cap N-terminal aa residues 21 to 41 interacted with p32. HEK293T cells were cotransfected with plasmids encoding the N-terminal aa residues 1 to 41, 1 to 20, or 21 to 41 fused with a GFP tag, along with Flag-p32; cell lysates were immunoprecipitated with anti-GFP antibody, followed by immunoblotting using indicated antibodies. (C) The Cap mutants were constructed with arginine residues at residues 24 to 26, 33, 35, and 37 replaced with alanine. (D) Mapping the crucial amino acids of Cap responsible for Cap/p32 interaction. HEK293T cells were cotransfected with Cap or Cap mutants 1, 2, and 3, along with Flag-p32 expression vectors, and the cell lysates were subjected to immunoprecipitation and immunoblotting using the indicated antibodies. (E) Confirmation by immunofluorescence assay of the crucial amino acids of PCV2 Cap that interact with p32. PK-15 cells were cotransfected with plasmids indicated in panel D and subjected to confocal microscopy observation at 36 h posttransfection. These results were confirmed in three independent experiments.
    Figure Legend Snippet: Cap binds to p32 via its N-terminal 24RRR26. (A) Cap fragment (aa 1 to 41) interacted with p32. HEK293T cells were cotransfected with plasmids encoding full-length Cap or deletion mutants fused with a GFP tag, along with Flag-p32; cell lysates were immunoprecipitated with an anti-GFP antibody and immunoblotted using indicated antibodies. (B) The Cap N-terminal aa residues 21 to 41 interacted with p32. HEK293T cells were cotransfected with plasmids encoding the N-terminal aa residues 1 to 41, 1 to 20, or 21 to 41 fused with a GFP tag, along with Flag-p32; cell lysates were immunoprecipitated with anti-GFP antibody, followed by immunoblotting using indicated antibodies. (C) The Cap mutants were constructed with arginine residues at residues 24 to 26, 33, 35, and 37 replaced with alanine. (D) Mapping the crucial amino acids of Cap responsible for Cap/p32 interaction. HEK293T cells were cotransfected with Cap or Cap mutants 1, 2, and 3, along with Flag-p32 expression vectors, and the cell lysates were subjected to immunoprecipitation and immunoblotting using the indicated antibodies. (E) Confirmation by immunofluorescence assay of the crucial amino acids of PCV2 Cap that interact with p32. PK-15 cells were cotransfected with plasmids indicated in panel D and subjected to confocal microscopy observation at 36 h posttransfection. These results were confirmed in three independent experiments.

    Techniques Used: Immunoprecipitation, Western Blot, Construct, Expressing, Immunofluorescence, Confocal Microscopy

    Model of PCV2 nuclear egress. PCV2 infection promotes the phosphorylation of PKC-δ via PLC-mediated signaling at the early infection phase, which is further amplified by JNK and ERK1/2 signaling at the late infection phase. In wild-type cells, Cap recruits p32 and phosphorylates PKC-δ (p-PKC-δ), and this leads to transport of p32 and p-PKC-δ into the nucleus. In the nucleus, p32, as an adaptor, further recruits p-PKC-δ and Cap to the nuclear membrane via binding with LBR to phosphorylate lamin A/C, which promotes the rearrangement of nuclear lamina and facilitates viral nuclear egress.
    Figure Legend Snippet: Model of PCV2 nuclear egress. PCV2 infection promotes the phosphorylation of PKC-δ via PLC-mediated signaling at the early infection phase, which is further amplified by JNK and ERK1/2 signaling at the late infection phase. In wild-type cells, Cap recruits p32 and phosphorylates PKC-δ (p-PKC-δ), and this leads to transport of p32 and p-PKC-δ into the nucleus. In the nucleus, p32, as an adaptor, further recruits p-PKC-δ and Cap to the nuclear membrane via binding with LBR to phosphorylate lamin A/C, which promotes the rearrangement of nuclear lamina and facilitates viral nuclear egress.

    Techniques Used: Infection, Amplification, Binding Assay

    gc1qr, human, mab 60.11  (Hycult Biotech)


    Bioz Verified Symbol Hycult Biotech is a verified supplier
    Bioz Manufacturer Symbol Hycult Biotech manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 90

    Structured Review

    Hycult Biotech gc1qr, human, mab 60.11
    Gc1qr, Human, Mab 60.11, supplied by Hycult Biotech, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/gc1qr, human, mab 60.11/product/Hycult Biotech
    Average 90 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    gc1qr, human, mab 60.11 - by Bioz Stars, 2024-05
    90/100 stars

    Images

    Similar Products

  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 90
    Hycult Biotech mouse monoclonal anti p32 antibody
    Mouse Monoclonal Anti P32 Antibody, supplied by Hycult Biotech, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/mouse monoclonal anti p32 antibody/product/Hycult Biotech
    Average 90 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    mouse monoclonal anti p32 antibody - by Bioz Stars, 2024-05
    90/100 stars
      Buy from Supplier

    90
    Hycult Biotech gc1qr, human, mab 60.11
    Gc1qr, Human, Mab 60.11, supplied by Hycult Biotech, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/gc1qr, human, mab 60.11/product/Hycult Biotech
    Average 90 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    gc1qr, human, mab 60.11 - by Bioz Stars, 2024-05
    90/100 stars
      Buy from Supplier

    Image Search Results