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rabbit monoclonal anti cpsf6  (Novus Biologicals)


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    Novus Biologicals rabbit monoclonal anti cpsf6
    Spinning disk confocal images of HEK293 SRRM2 HaloTag cells acquired 9 hr after VSV-G/HIV-1ΔEnvIN HA LAI (BRU) (MOI 10) infection occurred in the presence of Nevirapine (10 µM). Cells were previously transduced for 24 hr with CPSF6-mNeonGreen lentiviral vector (MOI 0.5) to detect CPSF6 (visualized in green). SRRM2 MLOs were detected using the TMR-HaloTag Ligand (in red) and the nuclei were stained with Hoechst (blue). Acquisitions were performed continuously for 27 min, which allowed us to see the independent formation of CPSF6 puncta, their migration towards the SRRM2 MLOs and the final fusion of CPSF6 puncta and SRRM2 MLOs.
    Rabbit Monoclonal Anti Cpsf6, supplied by Novus Biologicals, used in various techniques. Bioz Stars score: 94/100, based on 10 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/rabbit+anti+cpsf6+antibody/CPSF6+Antibody/pmc12774418-10-2-6
    Average 94 stars, based on 10 article reviews
    rabbit monoclonal anti cpsf6 - by Bioz Stars, 2026-09
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    Images

    1) Product Images from "Decoding the biogenesis of HIV-induced CPSF6 puncta and their fusion with nuclear speckles"

    Article Title: Decoding the biogenesis of HIV-induced CPSF6 puncta and their fusion with nuclear speckles

    Journal: eLife

    doi: 10.7554/eLife.103725

    Spinning disk confocal images of HEK293 SRRM2 HaloTag cells acquired 9 hr after VSV-G/HIV-1ΔEnvIN HA LAI (BRU) (MOI 10) infection occurred in the presence of Nevirapine (10 µM). Cells were previously transduced for 24 hr with CPSF6-mNeonGreen lentiviral vector (MOI 0.5) to detect CPSF6 (visualized in green). SRRM2 MLOs were detected using the TMR-HaloTag Ligand (in red) and the nuclei were stained with Hoechst (blue). Acquisitions were performed continuously for 27 min, which allowed us to see the independent formation of CPSF6 puncta, their migration towards the SRRM2 MLOs and the final fusion of CPSF6 puncta and SRRM2 MLOs.
    Figure Legend Snippet: Spinning disk confocal images of HEK293 SRRM2 HaloTag cells acquired 9 hr after VSV-G/HIV-1ΔEnvIN HA LAI (BRU) (MOI 10) infection occurred in the presence of Nevirapine (10 µM). Cells were previously transduced for 24 hr with CPSF6-mNeonGreen lentiviral vector (MOI 0.5) to detect CPSF6 (visualized in green). SRRM2 MLOs were detected using the TMR-HaloTag Ligand (in red) and the nuclei were stained with Hoechst (blue). Acquisitions were performed continuously for 27 min, which allowed us to see the independent formation of CPSF6 puncta, their migration towards the SRRM2 MLOs and the final fusion of CPSF6 puncta and SRRM2 MLOs.

    Techniques Used:

    Spinning disk confocal images of HEK293 SRRM2 HaloTag cells acquired 9 hr after VSV-G/HIV-1ΔEnvIN HA LAI (BRU) (MOI 10) infection occurred in the presence of Nevirapine (10 µM). Cells were previously transduced for 24 hr with CPSF6-mNeonGreen lentiviral vector (MOI 0.5) to detect CPSF6 (visualized in green). SRRM2 MLOs were detected using the TMR-HaloTag Ligand (in red) and the nuclei were stained with Hoechst (blue). Acquisitions were performed continuously for 27 min, which allowed us to see the independent formation of CPSF6 puncta, their migration towards the SRRM2 MLOs and the final fusion of CPSF6 puncta and SRRM2 MLOs.
    Figure Legend Snippet: Spinning disk confocal images of HEK293 SRRM2 HaloTag cells acquired 9 hr after VSV-G/HIV-1ΔEnvIN HA LAI (BRU) (MOI 10) infection occurred in the presence of Nevirapine (10 µM). Cells were previously transduced for 24 hr with CPSF6-mNeonGreen lentiviral vector (MOI 0.5) to detect CPSF6 (visualized in green). SRRM2 MLOs were detected using the TMR-HaloTag Ligand (in red) and the nuclei were stained with Hoechst (blue). Acquisitions were performed continuously for 27 min, which allowed us to see the independent formation of CPSF6 puncta, their migration towards the SRRM2 MLOs and the final fusion of CPSF6 puncta and SRRM2 MLOs.

    Techniques Used:

    Related Articles

    Staining:

    Article Title: The ability of TNPO3-depleted cells to inhibit HIV-1 infection requires CPSF6
    Article Snippet: The TNPO3 protein was stained using rabbit (Abcam cat# ab65016) or a mouse (cat# ab54353) anti-TNPO3 antibody. .. The CPSF6 protein was stained using a rabbit anti-CPSF6 antibody (Novus Biological cat# NB100-61596). .. The ASF/SF2 protein was stained using a mouse anti-ASF/SF2 antibody (Santa Cruz Biotechnology cat# cs-73026).



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    Figure 1. Downregulated <t>CPSF6</t> protein in macrophages upon viral infection (A) Notched boxplot of weighted mean 30 UTR length of THP-1 cells upon VSV (MOI ꞊1) infection at 0, 4, 8 and 12 h. ****p < 0.0001 with paired t test. Two biological repeats were performed with IVT-SAPAS. (B) Heatmap of mRNA expression levels of 30 end-processing factors. (C) Western blot and RT-qPCR analysis of CPSF6 protein and mRNA levels in THP-1 cells upon VSV (MOI ꞊1) infection at the indicated time points. b-actin was used as a loading control. Error bars indicate SD (n = 3 replicates). (D and E) Protein and mRNA expression levels of CPSF6 in C57BL/6 mouse PMs and BMDMs infected with VSV at different time points. Error bars indicate SD (n = 3 replicates). (F and G) Protein and mRNA expression levels of CPSF6 in THP-1 cells stimulated with poly(dA:dT) and poly(I:C). All RT-qPCR experiments were performed with three biological replicates, and b-actin was used as an internal control. Error bars indicate SD (n = 3 replicates).
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    Image Search Results


    Spinning disk confocal images of HEK293 SRRM2 HaloTag cells acquired 9 hr after VSV-G/HIV-1ΔEnvIN HA LAI (BRU) (MOI 10) infection occurred in the presence of Nevirapine (10 µM). Cells were previously transduced for 24 hr with CPSF6-mNeonGreen lentiviral vector (MOI 0.5) to detect CPSF6 (visualized in green). SRRM2 MLOs were detected using the TMR-HaloTag Ligand (in red) and the nuclei were stained with Hoechst (blue). Acquisitions were performed continuously for 27 min, which allowed us to see the independent formation of CPSF6 puncta, their migration towards the SRRM2 MLOs and the final fusion of CPSF6 puncta and SRRM2 MLOs.

    Journal: eLife

    Article Title: Decoding the biogenesis of HIV-induced CPSF6 puncta and their fusion with nuclear speckles

    doi: 10.7554/eLife.103725

    Figure Lengend Snippet: Spinning disk confocal images of HEK293 SRRM2 HaloTag cells acquired 9 hr after VSV-G/HIV-1ΔEnvIN HA LAI (BRU) (MOI 10) infection occurred in the presence of Nevirapine (10 µM). Cells were previously transduced for 24 hr with CPSF6-mNeonGreen lentiviral vector (MOI 0.5) to detect CPSF6 (visualized in green). SRRM2 MLOs were detected using the TMR-HaloTag Ligand (in red) and the nuclei were stained with Hoechst (blue). Acquisitions were performed continuously for 27 min, which allowed us to see the independent formation of CPSF6 puncta, their migration towards the SRRM2 MLOs and the final fusion of CPSF6 puncta and SRRM2 MLOs.

    Article Snippet: Antibody , Rabbit monoclonal anti-CPSF6 , Novus Biologicals , #NBP1-85676 , IF 1:400 WB 1:500.

    Techniques:

    Spinning disk confocal images of HEK293 SRRM2 HaloTag cells acquired 9 hr after VSV-G/HIV-1ΔEnvIN HA LAI (BRU) (MOI 10) infection occurred in the presence of Nevirapine (10 µM). Cells were previously transduced for 24 hr with CPSF6-mNeonGreen lentiviral vector (MOI 0.5) to detect CPSF6 (visualized in green). SRRM2 MLOs were detected using the TMR-HaloTag Ligand (in red) and the nuclei were stained with Hoechst (blue). Acquisitions were performed continuously for 27 min, which allowed us to see the independent formation of CPSF6 puncta, their migration towards the SRRM2 MLOs and the final fusion of CPSF6 puncta and SRRM2 MLOs.

    Journal: eLife

    Article Title: Decoding the biogenesis of HIV-induced CPSF6 puncta and their fusion with nuclear speckles

    doi: 10.7554/eLife.103725

    Figure Lengend Snippet: Spinning disk confocal images of HEK293 SRRM2 HaloTag cells acquired 9 hr after VSV-G/HIV-1ΔEnvIN HA LAI (BRU) (MOI 10) infection occurred in the presence of Nevirapine (10 µM). Cells were previously transduced for 24 hr with CPSF6-mNeonGreen lentiviral vector (MOI 0.5) to detect CPSF6 (visualized in green). SRRM2 MLOs were detected using the TMR-HaloTag Ligand (in red) and the nuclei were stained with Hoechst (blue). Acquisitions were performed continuously for 27 min, which allowed us to see the independent formation of CPSF6 puncta, their migration towards the SRRM2 MLOs and the final fusion of CPSF6 puncta and SRRM2 MLOs.

    Article Snippet: Antibody , Rabbit monoclonal anti-CPSF6 , Novus Biologicals , #NBP1-85676 , IF 1:400 WB 1:500.

    Techniques:

    Figure 1. Downregulated CPSF6 protein in macrophages upon viral infection (A) Notched boxplot of weighted mean 30 UTR length of THP-1 cells upon VSV (MOI ꞊1) infection at 0, 4, 8 and 12 h. ****p < 0.0001 with paired t test. Two biological repeats were performed with IVT-SAPAS. (B) Heatmap of mRNA expression levels of 30 end-processing factors. (C) Western blot and RT-qPCR analysis of CPSF6 protein and mRNA levels in THP-1 cells upon VSV (MOI ꞊1) infection at the indicated time points. b-actin was used as a loading control. Error bars indicate SD (n = 3 replicates). (D and E) Protein and mRNA expression levels of CPSF6 in C57BL/6 mouse PMs and BMDMs infected with VSV at different time points. Error bars indicate SD (n = 3 replicates). (F and G) Protein and mRNA expression levels of CPSF6 in THP-1 cells stimulated with poly(dA:dT) and poly(I:C). All RT-qPCR experiments were performed with three biological replicates, and b-actin was used as an internal control. Error bars indicate SD (n = 3 replicates).

    Journal: Cell reports

    Article Title: E3 ligase SYVN1-mediated polyubiquitination of CPSF6 promotes alternative polyadenylation and antivirus effects of macrophages.

    doi: 10.1016/j.celrep.2025.115276

    Figure Lengend Snippet: Figure 1. Downregulated CPSF6 protein in macrophages upon viral infection (A) Notched boxplot of weighted mean 30 UTR length of THP-1 cells upon VSV (MOI ꞊1) infection at 0, 4, 8 and 12 h. ****p < 0.0001 with paired t test. Two biological repeats were performed with IVT-SAPAS. (B) Heatmap of mRNA expression levels of 30 end-processing factors. (C) Western blot and RT-qPCR analysis of CPSF6 protein and mRNA levels in THP-1 cells upon VSV (MOI ꞊1) infection at the indicated time points. b-actin was used as a loading control. Error bars indicate SD (n = 3 replicates). (D and E) Protein and mRNA expression levels of CPSF6 in C57BL/6 mouse PMs and BMDMs infected with VSV at different time points. Error bars indicate SD (n = 3 replicates). (F and G) Protein and mRNA expression levels of CPSF6 in THP-1 cells stimulated with poly(dA:dT) and poly(I:C). All RT-qPCR experiments were performed with three biological replicates, and b-actin was used as an internal control. Error bars indicate SD (n = 3 replicates).

    Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Rabbit polyclonal anti-CPSF6 Novus Biologicals Cat# NBP1-85676; RRID: AB_11004635 anti-b-actin FUDE Biologicals Cat# FD0060; RRID: AB_2923199 Mouse polyclonal anti-CPSF5 Proteintech Cat# 10322-1-AP; RRID: AB_2251496 Rabbit polyclonal anti-CPSF7 Proteintech Cat# 55195-1-AP; RRID: AB_10858797 Recombinant Anti-SYVN1/HRD1 antibody abcam Cat# ab170901; RRID: AB_2833021 Rabbit polyclonal anti-HRD1 Novus Cat# NB100-2526; RRID: AB_608700 Ubiquitin (P37) Antibody Cell Signaling Technology Cat# 58395S; RRID: AB_3075532 Ubiquitin Antibody Santa Cruz Biotechnology Cat# sc-271289; RRID: AB_10611436 Rabbit Anti-HA Tag RB0411 Polyclonal Antibody abcepta Cat# AP1012a; RRID: AB_352510 Mouse monoclonal anti-Myc Sigma Cat# M4439; RRID: AB_439694 Monoclonal ANTI-FLAG M2 antibody produced in mouse Sigma Cat# F1804; RRID: AB_262044 K48-linkage Specific Polyubiquitin (D9D5) Rabbit mAb Cell Signaling Technology Cat# 8081; RRID: AB_10859893 K63-linkage Specific Polyubiquitin (D7A11) Rabbit mAb Cell Signaling Technology Cat# 5621; RRID: AB_10827985 Anti-rabbit IgG Cell Signaling Technology Cat# 7074; RRID: AB_2099233 Anti-mouse IgG HRP-linked Antibody Cell Signaling Technology Cat# 7076; RRID: AB_330924 Goat anti-Rabbit IgG (H + L) CrossAdsorbed Secondary Antibody, Alexa FluorTM 568 Invitrogen Cat# A-11011; RRID: AB_143157 Goat anti-rabbit IgG (H + L) CrossAdsorbed Secondary Antibody, Alexa FluorTM 488 Invitrogen Cat# A-11008; RRID: AB_143165 anti-Lamin B1 Cell Signaling Technology Cat# 13435; RRID: AB_2737428 MyD88 (D80F5) Rabbit mAb Cell Signaling Technology Cat# 4283; RRID: AB_10547882 MAVS (D5A9E) Rabbit mAb Cell Signaling Technology Cat# 24930; RRID: AB_2798889 RIG-I (D14G6) Rabbit mAb Cell Signaling Technology Cat# 3743; RRID: AB_2269233 TRIF Antibody Cell Signaling Technology Cat# 4596; RRID: AB_2256555 MDA5 Recombinant Rabbit Monoclonal Antibody (33H12L34) Thermo Fisher Scientific Cat# 700360; RRID: AB_2532316 (Continued on next page) Cell Reports 44, 115276, February 25, 2025 15

    Techniques: Infection, Expressing, Western Blot, Quantitative RT-PCR, Control

    Figure 2. CPSF6 is degraded through the ubiquitination-proteasome pathway by the E3 ligase SYVN1 (A) Western blot analysis of CPSF6 protein expression in THP-1 cells with treatment of MG132 (10 mM) or DMSO. The cells were infected with VSV-EGFP (MOI ꞊1) for 4 and 8 h before sample collection. (B) Elevated ubiquitination level of CPSF6 upon VSV infection. THP-1 cells were treated with MG132 or DMSO and then infected with VSV- EGFP for 4 and 8 h, followed by IP with anti- CPSF6 and western blot with anti-Ub and anti- CPSF6. (C) Knockdown of SYVN1 attenuates the ubiq- uitination of CPSF6 upon VSV infection. THP-1 cells were transfected with SYVN1 siRNA for 48 h and infected with VSV-EGFP for 8 h (n = 3 replicates). (D) The SYVN1 inhibitor represses the ubiquiti- nation of CPSF6. THP-1 cells were treated with SYVN1 inhibitor LS-102 (10 mM) and MG132 for 8 h and infected with VSV-EGFP for 8 h, followed by IP with anti-CPSF6 and western blot with anti- Ub and anti-CPSF6. Homologous IgG was used as a negative control.

    Journal: Cell reports

    Article Title: E3 ligase SYVN1-mediated polyubiquitination of CPSF6 promotes alternative polyadenylation and antivirus effects of macrophages.

    doi: 10.1016/j.celrep.2025.115276

    Figure Lengend Snippet: Figure 2. CPSF6 is degraded through the ubiquitination-proteasome pathway by the E3 ligase SYVN1 (A) Western blot analysis of CPSF6 protein expression in THP-1 cells with treatment of MG132 (10 mM) or DMSO. The cells were infected with VSV-EGFP (MOI ꞊1) for 4 and 8 h before sample collection. (B) Elevated ubiquitination level of CPSF6 upon VSV infection. THP-1 cells were treated with MG132 or DMSO and then infected with VSV- EGFP for 4 and 8 h, followed by IP with anti- CPSF6 and western blot with anti-Ub and anti- CPSF6. (C) Knockdown of SYVN1 attenuates the ubiq- uitination of CPSF6 upon VSV infection. THP-1 cells were transfected with SYVN1 siRNA for 48 h and infected with VSV-EGFP for 8 h (n = 3 replicates). (D) The SYVN1 inhibitor represses the ubiquiti- nation of CPSF6. THP-1 cells were treated with SYVN1 inhibitor LS-102 (10 mM) and MG132 for 8 h and infected with VSV-EGFP for 8 h, followed by IP with anti-CPSF6 and western blot with anti- Ub and anti-CPSF6. Homologous IgG was used as a negative control.

    Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Rabbit polyclonal anti-CPSF6 Novus Biologicals Cat# NBP1-85676; RRID: AB_11004635 anti-b-actin FUDE Biologicals Cat# FD0060; RRID: AB_2923199 Mouse polyclonal anti-CPSF5 Proteintech Cat# 10322-1-AP; RRID: AB_2251496 Rabbit polyclonal anti-CPSF7 Proteintech Cat# 55195-1-AP; RRID: AB_10858797 Recombinant Anti-SYVN1/HRD1 antibody abcam Cat# ab170901; RRID: AB_2833021 Rabbit polyclonal anti-HRD1 Novus Cat# NB100-2526; RRID: AB_608700 Ubiquitin (P37) Antibody Cell Signaling Technology Cat# 58395S; RRID: AB_3075532 Ubiquitin Antibody Santa Cruz Biotechnology Cat# sc-271289; RRID: AB_10611436 Rabbit Anti-HA Tag RB0411 Polyclonal Antibody abcepta Cat# AP1012a; RRID: AB_352510 Mouse monoclonal anti-Myc Sigma Cat# M4439; RRID: AB_439694 Monoclonal ANTI-FLAG M2 antibody produced in mouse Sigma Cat# F1804; RRID: AB_262044 K48-linkage Specific Polyubiquitin (D9D5) Rabbit mAb Cell Signaling Technology Cat# 8081; RRID: AB_10859893 K63-linkage Specific Polyubiquitin (D7A11) Rabbit mAb Cell Signaling Technology Cat# 5621; RRID: AB_10827985 Anti-rabbit IgG Cell Signaling Technology Cat# 7074; RRID: AB_2099233 Anti-mouse IgG HRP-linked Antibody Cell Signaling Technology Cat# 7076; RRID: AB_330924 Goat anti-Rabbit IgG (H + L) CrossAdsorbed Secondary Antibody, Alexa FluorTM 568 Invitrogen Cat# A-11011; RRID: AB_143157 Goat anti-rabbit IgG (H + L) CrossAdsorbed Secondary Antibody, Alexa FluorTM 488 Invitrogen Cat# A-11008; RRID: AB_143165 anti-Lamin B1 Cell Signaling Technology Cat# 13435; RRID: AB_2737428 MyD88 (D80F5) Rabbit mAb Cell Signaling Technology Cat# 4283; RRID: AB_10547882 MAVS (D5A9E) Rabbit mAb Cell Signaling Technology Cat# 24930; RRID: AB_2798889 RIG-I (D14G6) Rabbit mAb Cell Signaling Technology Cat# 3743; RRID: AB_2269233 TRIF Antibody Cell Signaling Technology Cat# 4596; RRID: AB_2256555 MDA5 Recombinant Rabbit Monoclonal Antibody (33H12L34) Thermo Fisher Scientific Cat# 700360; RRID: AB_2532316 (Continued on next page) Cell Reports 44, 115276, February 25, 2025 15

    Techniques: Ubiquitin Proteomics, Western Blot, Expressing, Infection, Knockdown, Transfection, Negative Control

    Figure 3. The interaction of SYVN1 with CPSF6 (A and B) Endogenous CPSF6 and SYVN1 can pull down each other. THP-1 cells were treated with MG132 (10 mM) and VSV infection for 8 h, and coIP was performed with anti-CPSF6 and anti-SYVN1. (C and D) Overexpressed Myc-CPSF6 and FLAG-SYVN1 can pull down each other. Myc-CPSF6 and FLAG-SYVN1 were co-transfected into HEK293T cells for 48 h, and coIP was performed with anti-Myc and anti-FLAG, respectively. (E) More SYVN1 can be pulled down by CPSF6 along with the infection time of VSV-EGFP. THP-1 cells were treated with MG132 (10 mM) and infected with VSV- EGFP. At different time points of infection, samples were collected, and coIP was performed with anti-CPSF6. (F) Schematic of fragments of human CPSF6 fused with the Myc tag: WT, RNA recognition motif (RRM) domain, proline-rich domain (PRD), and arginine/serine- like domain (RSLD). (G) Myc-CPSF6-RRM can pull down FLAG-SYVN1. HEK293T cells were co-transfected with FLAG-SYVN1 and Myc-CPSF6 fragments for 48 h, and coIP with anti-Myc was performed. Homologous IgG was used as a negative control.

    Journal: Cell reports

    Article Title: E3 ligase SYVN1-mediated polyubiquitination of CPSF6 promotes alternative polyadenylation and antivirus effects of macrophages.

    doi: 10.1016/j.celrep.2025.115276

    Figure Lengend Snippet: Figure 3. The interaction of SYVN1 with CPSF6 (A and B) Endogenous CPSF6 and SYVN1 can pull down each other. THP-1 cells were treated with MG132 (10 mM) and VSV infection for 8 h, and coIP was performed with anti-CPSF6 and anti-SYVN1. (C and D) Overexpressed Myc-CPSF6 and FLAG-SYVN1 can pull down each other. Myc-CPSF6 and FLAG-SYVN1 were co-transfected into HEK293T cells for 48 h, and coIP was performed with anti-Myc and anti-FLAG, respectively. (E) More SYVN1 can be pulled down by CPSF6 along with the infection time of VSV-EGFP. THP-1 cells were treated with MG132 (10 mM) and infected with VSV- EGFP. At different time points of infection, samples were collected, and coIP was performed with anti-CPSF6. (F) Schematic of fragments of human CPSF6 fused with the Myc tag: WT, RNA recognition motif (RRM) domain, proline-rich domain (PRD), and arginine/serine- like domain (RSLD). (G) Myc-CPSF6-RRM can pull down FLAG-SYVN1. HEK293T cells were co-transfected with FLAG-SYVN1 and Myc-CPSF6 fragments for 48 h, and coIP with anti-Myc was performed. Homologous IgG was used as a negative control.

    Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Rabbit polyclonal anti-CPSF6 Novus Biologicals Cat# NBP1-85676; RRID: AB_11004635 anti-b-actin FUDE Biologicals Cat# FD0060; RRID: AB_2923199 Mouse polyclonal anti-CPSF5 Proteintech Cat# 10322-1-AP; RRID: AB_2251496 Rabbit polyclonal anti-CPSF7 Proteintech Cat# 55195-1-AP; RRID: AB_10858797 Recombinant Anti-SYVN1/HRD1 antibody abcam Cat# ab170901; RRID: AB_2833021 Rabbit polyclonal anti-HRD1 Novus Cat# NB100-2526; RRID: AB_608700 Ubiquitin (P37) Antibody Cell Signaling Technology Cat# 58395S; RRID: AB_3075532 Ubiquitin Antibody Santa Cruz Biotechnology Cat# sc-271289; RRID: AB_10611436 Rabbit Anti-HA Tag RB0411 Polyclonal Antibody abcepta Cat# AP1012a; RRID: AB_352510 Mouse monoclonal anti-Myc Sigma Cat# M4439; RRID: AB_439694 Monoclonal ANTI-FLAG M2 antibody produced in mouse Sigma Cat# F1804; RRID: AB_262044 K48-linkage Specific Polyubiquitin (D9D5) Rabbit mAb Cell Signaling Technology Cat# 8081; RRID: AB_10859893 K63-linkage Specific Polyubiquitin (D7A11) Rabbit mAb Cell Signaling Technology Cat# 5621; RRID: AB_10827985 Anti-rabbit IgG Cell Signaling Technology Cat# 7074; RRID: AB_2099233 Anti-mouse IgG HRP-linked Antibody Cell Signaling Technology Cat# 7076; RRID: AB_330924 Goat anti-Rabbit IgG (H + L) CrossAdsorbed Secondary Antibody, Alexa FluorTM 568 Invitrogen Cat# A-11011; RRID: AB_143157 Goat anti-rabbit IgG (H + L) CrossAdsorbed Secondary Antibody, Alexa FluorTM 488 Invitrogen Cat# A-11008; RRID: AB_143165 anti-Lamin B1 Cell Signaling Technology Cat# 13435; RRID: AB_2737428 MyD88 (D80F5) Rabbit mAb Cell Signaling Technology Cat# 4283; RRID: AB_10547882 MAVS (D5A9E) Rabbit mAb Cell Signaling Technology Cat# 24930; RRID: AB_2798889 RIG-I (D14G6) Rabbit mAb Cell Signaling Technology Cat# 3743; RRID: AB_2269233 TRIF Antibody Cell Signaling Technology Cat# 4596; RRID: AB_2256555 MDA5 Recombinant Rabbit Monoclonal Antibody (33H12L34) Thermo Fisher Scientific Cat# 700360; RRID: AB_2532316 (Continued on next page) Cell Reports 44, 115276, February 25, 2025 15

    Techniques: Infection, Transfection, Negative Control

    Figure 4. SYVN1 catalyzes K48-linked ubiquitination of CPSF6 (A) Schematics of the HA-Ub mutant with single lysine only and the FLAG-SYVN1-C329S mutant. (B) Ubiquitination effects of Ub mutants on Myc-CPSF6 in HEK293T cells. The cells were co-transfected with Myc-CPSF6, FLAG-SYVN1, and HA-Ub (WT) or HA- Ub mutants for 48 h and infected with VSV, and IP was performed with anti-Myc following western blotting with anti-HA. (C) Ubiquitination effects of Ub mutants on endogenous CPSF6 in HEK293T cells. The cells were transfected with HA-Ub (WT) or HA-Ub mutants for 48 h, treated with MG132 (10 mM) or DMSO for 8 h, and infected with VSV, followed by IP with anti-CPSF6 and western blotting with anti-HA. (D) K48-linked ubiquitination of Myc-CPSF6 by FLAG-SYVN1 in HEK293T cells. The cells were co-transfected with Myc-CPSF6, FLAG-SYVN1, and HA-Ub (WT) and infected with VSV, followed by IP with anti-Myc and western blotting with anti-HA, anti-K48, and anti-K63. (E) The reduced effect of the SYVN1 C329S mutant on ubiquitination of Myc-CPSF6 in HEK293T cells. FLAG-SYVN1 and FLAG-SYVN1-C329S were co- transfected with Myc-CPSF6 and HA-K48, respectively, in HEK293T cells, followed by VSV infection and IP with anti-Myc and western blotting with anti-HA. Homologous IgG was used as a negative control.

    Journal: Cell reports

    Article Title: E3 ligase SYVN1-mediated polyubiquitination of CPSF6 promotes alternative polyadenylation and antivirus effects of macrophages.

    doi: 10.1016/j.celrep.2025.115276

    Figure Lengend Snippet: Figure 4. SYVN1 catalyzes K48-linked ubiquitination of CPSF6 (A) Schematics of the HA-Ub mutant with single lysine only and the FLAG-SYVN1-C329S mutant. (B) Ubiquitination effects of Ub mutants on Myc-CPSF6 in HEK293T cells. The cells were co-transfected with Myc-CPSF6, FLAG-SYVN1, and HA-Ub (WT) or HA- Ub mutants for 48 h and infected with VSV, and IP was performed with anti-Myc following western blotting with anti-HA. (C) Ubiquitination effects of Ub mutants on endogenous CPSF6 in HEK293T cells. The cells were transfected with HA-Ub (WT) or HA-Ub mutants for 48 h, treated with MG132 (10 mM) or DMSO for 8 h, and infected with VSV, followed by IP with anti-CPSF6 and western blotting with anti-HA. (D) K48-linked ubiquitination of Myc-CPSF6 by FLAG-SYVN1 in HEK293T cells. The cells were co-transfected with Myc-CPSF6, FLAG-SYVN1, and HA-Ub (WT) and infected with VSV, followed by IP with anti-Myc and western blotting with anti-HA, anti-K48, and anti-K63. (E) The reduced effect of the SYVN1 C329S mutant on ubiquitination of Myc-CPSF6 in HEK293T cells. FLAG-SYVN1 and FLAG-SYVN1-C329S were co- transfected with Myc-CPSF6 and HA-K48, respectively, in HEK293T cells, followed by VSV infection and IP with anti-Myc and western blotting with anti-HA. Homologous IgG was used as a negative control.

    Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Rabbit polyclonal anti-CPSF6 Novus Biologicals Cat# NBP1-85676; RRID: AB_11004635 anti-b-actin FUDE Biologicals Cat# FD0060; RRID: AB_2923199 Mouse polyclonal anti-CPSF5 Proteintech Cat# 10322-1-AP; RRID: AB_2251496 Rabbit polyclonal anti-CPSF7 Proteintech Cat# 55195-1-AP; RRID: AB_10858797 Recombinant Anti-SYVN1/HRD1 antibody abcam Cat# ab170901; RRID: AB_2833021 Rabbit polyclonal anti-HRD1 Novus Cat# NB100-2526; RRID: AB_608700 Ubiquitin (P37) Antibody Cell Signaling Technology Cat# 58395S; RRID: AB_3075532 Ubiquitin Antibody Santa Cruz Biotechnology Cat# sc-271289; RRID: AB_10611436 Rabbit Anti-HA Tag RB0411 Polyclonal Antibody abcepta Cat# AP1012a; RRID: AB_352510 Mouse monoclonal anti-Myc Sigma Cat# M4439; RRID: AB_439694 Monoclonal ANTI-FLAG M2 antibody produced in mouse Sigma Cat# F1804; RRID: AB_262044 K48-linkage Specific Polyubiquitin (D9D5) Rabbit mAb Cell Signaling Technology Cat# 8081; RRID: AB_10859893 K63-linkage Specific Polyubiquitin (D7A11) Rabbit mAb Cell Signaling Technology Cat# 5621; RRID: AB_10827985 Anti-rabbit IgG Cell Signaling Technology Cat# 7074; RRID: AB_2099233 Anti-mouse IgG HRP-linked Antibody Cell Signaling Technology Cat# 7076; RRID: AB_330924 Goat anti-Rabbit IgG (H + L) CrossAdsorbed Secondary Antibody, Alexa FluorTM 568 Invitrogen Cat# A-11011; RRID: AB_143157 Goat anti-rabbit IgG (H + L) CrossAdsorbed Secondary Antibody, Alexa FluorTM 488 Invitrogen Cat# A-11008; RRID: AB_143165 anti-Lamin B1 Cell Signaling Technology Cat# 13435; RRID: AB_2737428 MyD88 (D80F5) Rabbit mAb Cell Signaling Technology Cat# 4283; RRID: AB_10547882 MAVS (D5A9E) Rabbit mAb Cell Signaling Technology Cat# 24930; RRID: AB_2798889 RIG-I (D14G6) Rabbit mAb Cell Signaling Technology Cat# 3743; RRID: AB_2269233 TRIF Antibody Cell Signaling Technology Cat# 4596; RRID: AB_2256555 MDA5 Recombinant Rabbit Monoclonal Antibody (33H12L34) Thermo Fisher Scientific Cat# 700360; RRID: AB_2532316 (Continued on next page) Cell Reports 44, 115276, February 25, 2025 15

    Techniques: Ubiquitin Proteomics, Mutagenesis, Transfection, Infection, Western Blot, Negative Control

    Figure 5. SYVN1 mediates K48-linked ubiquitination of CPSF6 at the sites of K161 and K185 (A) The potential ubiquitination sites (K79, K161, and K185) of CPSF6 predicted by UbiBrowser 2.0. (B) The effects of mutated potential ubiquitination sites on CPSF6 degradation mediated by SYVN1. The single-, double-, and triple-sites mutants of Myc-CPSF6 were co-transfected with FLAG-SYVN1 and HA-K48-Ub in HEK293T cells, followed by VSV infection and western blot analysis with the indicated antibodies.

    Journal: Cell reports

    Article Title: E3 ligase SYVN1-mediated polyubiquitination of CPSF6 promotes alternative polyadenylation and antivirus effects of macrophages.

    doi: 10.1016/j.celrep.2025.115276

    Figure Lengend Snippet: Figure 5. SYVN1 mediates K48-linked ubiquitination of CPSF6 at the sites of K161 and K185 (A) The potential ubiquitination sites (K79, K161, and K185) of CPSF6 predicted by UbiBrowser 2.0. (B) The effects of mutated potential ubiquitination sites on CPSF6 degradation mediated by SYVN1. The single-, double-, and triple-sites mutants of Myc-CPSF6 were co-transfected with FLAG-SYVN1 and HA-K48-Ub in HEK293T cells, followed by VSV infection and western blot analysis with the indicated antibodies.

    Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Rabbit polyclonal anti-CPSF6 Novus Biologicals Cat# NBP1-85676; RRID: AB_11004635 anti-b-actin FUDE Biologicals Cat# FD0060; RRID: AB_2923199 Mouse polyclonal anti-CPSF5 Proteintech Cat# 10322-1-AP; RRID: AB_2251496 Rabbit polyclonal anti-CPSF7 Proteintech Cat# 55195-1-AP; RRID: AB_10858797 Recombinant Anti-SYVN1/HRD1 antibody abcam Cat# ab170901; RRID: AB_2833021 Rabbit polyclonal anti-HRD1 Novus Cat# NB100-2526; RRID: AB_608700 Ubiquitin (P37) Antibody Cell Signaling Technology Cat# 58395S; RRID: AB_3075532 Ubiquitin Antibody Santa Cruz Biotechnology Cat# sc-271289; RRID: AB_10611436 Rabbit Anti-HA Tag RB0411 Polyclonal Antibody abcepta Cat# AP1012a; RRID: AB_352510 Mouse monoclonal anti-Myc Sigma Cat# M4439; RRID: AB_439694 Monoclonal ANTI-FLAG M2 antibody produced in mouse Sigma Cat# F1804; RRID: AB_262044 K48-linkage Specific Polyubiquitin (D9D5) Rabbit mAb Cell Signaling Technology Cat# 8081; RRID: AB_10859893 K63-linkage Specific Polyubiquitin (D7A11) Rabbit mAb Cell Signaling Technology Cat# 5621; RRID: AB_10827985 Anti-rabbit IgG Cell Signaling Technology Cat# 7074; RRID: AB_2099233 Anti-mouse IgG HRP-linked Antibody Cell Signaling Technology Cat# 7076; RRID: AB_330924 Goat anti-Rabbit IgG (H + L) CrossAdsorbed Secondary Antibody, Alexa FluorTM 568 Invitrogen Cat# A-11011; RRID: AB_143157 Goat anti-rabbit IgG (H + L) CrossAdsorbed Secondary Antibody, Alexa FluorTM 488 Invitrogen Cat# A-11008; RRID: AB_143165 anti-Lamin B1 Cell Signaling Technology Cat# 13435; RRID: AB_2737428 MyD88 (D80F5) Rabbit mAb Cell Signaling Technology Cat# 4283; RRID: AB_10547882 MAVS (D5A9E) Rabbit mAb Cell Signaling Technology Cat# 24930; RRID: AB_2798889 RIG-I (D14G6) Rabbit mAb Cell Signaling Technology Cat# 3743; RRID: AB_2269233 TRIF Antibody Cell Signaling Technology Cat# 4596; RRID: AB_2256555 MDA5 Recombinant Rabbit Monoclonal Antibody (33H12L34) Thermo Fisher Scientific Cat# 700360; RRID: AB_2532316 (Continued on next page) Cell Reports 44, 115276, February 25, 2025 15

    Techniques: Ubiquitin Proteomics, Transfection, Infection, Western Blot

    Figure 6. Anti-virus effect of SYVN1 through ubiquitination of CPSF6 (A) The effect of CPSF6 ubiquitination site mutation on VSV replication. Left: RT-qPCR analysis of VSV-G in L929 cells with stably expressed WT and mutants of CPSF6 at 24 h of VSV-EGFP infection. Error bars indicate SD (n = 3 replicates). Right: western blot analysis of VSV-G protein at 24 and 36 h of infection. ns, not significant; **p < 0.01, ***p < 0.001, ****p < 0.0001 with t test and Bonferroni correction. (B) RT-qPCR analysis of the immune-related genes in L929 cells with stable expression of the WT and mutants of CPSF6 at 24 h of VSV-EGFP infection. Error bars indicate SD (n = 3 replicates). **p < 0.01, ***p < 0.001 with t test and Bonferroni correction.

    Journal: Cell reports

    Article Title: E3 ligase SYVN1-mediated polyubiquitination of CPSF6 promotes alternative polyadenylation and antivirus effects of macrophages.

    doi: 10.1016/j.celrep.2025.115276

    Figure Lengend Snippet: Figure 6. Anti-virus effect of SYVN1 through ubiquitination of CPSF6 (A) The effect of CPSF6 ubiquitination site mutation on VSV replication. Left: RT-qPCR analysis of VSV-G in L929 cells with stably expressed WT and mutants of CPSF6 at 24 h of VSV-EGFP infection. Error bars indicate SD (n = 3 replicates). Right: western blot analysis of VSV-G protein at 24 and 36 h of infection. ns, not significant; **p < 0.01, ***p < 0.001, ****p < 0.0001 with t test and Bonferroni correction. (B) RT-qPCR analysis of the immune-related genes in L929 cells with stable expression of the WT and mutants of CPSF6 at 24 h of VSV-EGFP infection. Error bars indicate SD (n = 3 replicates). **p < 0.01, ***p < 0.001 with t test and Bonferroni correction.

    Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Rabbit polyclonal anti-CPSF6 Novus Biologicals Cat# NBP1-85676; RRID: AB_11004635 anti-b-actin FUDE Biologicals Cat# FD0060; RRID: AB_2923199 Mouse polyclonal anti-CPSF5 Proteintech Cat# 10322-1-AP; RRID: AB_2251496 Rabbit polyclonal anti-CPSF7 Proteintech Cat# 55195-1-AP; RRID: AB_10858797 Recombinant Anti-SYVN1/HRD1 antibody abcam Cat# ab170901; RRID: AB_2833021 Rabbit polyclonal anti-HRD1 Novus Cat# NB100-2526; RRID: AB_608700 Ubiquitin (P37) Antibody Cell Signaling Technology Cat# 58395S; RRID: AB_3075532 Ubiquitin Antibody Santa Cruz Biotechnology Cat# sc-271289; RRID: AB_10611436 Rabbit Anti-HA Tag RB0411 Polyclonal Antibody abcepta Cat# AP1012a; RRID: AB_352510 Mouse monoclonal anti-Myc Sigma Cat# M4439; RRID: AB_439694 Monoclonal ANTI-FLAG M2 antibody produced in mouse Sigma Cat# F1804; RRID: AB_262044 K48-linkage Specific Polyubiquitin (D9D5) Rabbit mAb Cell Signaling Technology Cat# 8081; RRID: AB_10859893 K63-linkage Specific Polyubiquitin (D7A11) Rabbit mAb Cell Signaling Technology Cat# 5621; RRID: AB_10827985 Anti-rabbit IgG Cell Signaling Technology Cat# 7074; RRID: AB_2099233 Anti-mouse IgG HRP-linked Antibody Cell Signaling Technology Cat# 7076; RRID: AB_330924 Goat anti-Rabbit IgG (H + L) CrossAdsorbed Secondary Antibody, Alexa FluorTM 568 Invitrogen Cat# A-11011; RRID: AB_143157 Goat anti-rabbit IgG (H + L) CrossAdsorbed Secondary Antibody, Alexa FluorTM 488 Invitrogen Cat# A-11008; RRID: AB_143165 anti-Lamin B1 Cell Signaling Technology Cat# 13435; RRID: AB_2737428 MyD88 (D80F5) Rabbit mAb Cell Signaling Technology Cat# 4283; RRID: AB_10547882 MAVS (D5A9E) Rabbit mAb Cell Signaling Technology Cat# 24930; RRID: AB_2798889 RIG-I (D14G6) Rabbit mAb Cell Signaling Technology Cat# 3743; RRID: AB_2269233 TRIF Antibody Cell Signaling Technology Cat# 4596; RRID: AB_2256555 MDA5 Recombinant Rabbit Monoclonal Antibody (33H12L34) Thermo Fisher Scientific Cat# 700360; RRID: AB_2532316 (Continued on next page) Cell Reports 44, 115276, February 25, 2025 15

    Techniques: Virus, Ubiquitin Proteomics, Mutagenesis, Quantitative RT-PCR, Stable Transfection, Infection, Western Blot, Expressing

    KEY RESOURCES TABLE

    Journal: Cell reports

    Article Title: CPSF1 inhibition promotes widespread use of intergenic polyadenylation sites and impairs glycolysis in prostate cancer cells

    doi: 10.1016/j.celrep.2024.115211

    Figure Lengend Snippet: KEY RESOURCES TABLE

    Article Snippet: Antibodies were purchased from Santa Cruz for western blot detection of CPSF1, (G-10, sc-166281, 1:100) and tubulin (B-5-1-2, sc-23948, 1:3000), Proteintech for western blot detection of NUDT21 (66335-1-Ig, 1:500) and PCF11 (23540-1-AP, 1:500), Fortis Life Sciences for western blot detection of CPSF6 (A301-356A, 1:1,000) and Origene for western blot detection of GPI (TA501171, 1:500).

    Techniques: Recombinant, shRNA, Over Expression, Plasmid Preparation, Cloning, Virus, Software