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rabbit polyclonal anti cpsf6 (Bethyl)

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Bethyl rabbit polyclonal anti cpsf6
Rabbit Polyclonal Anti Cpsf6, supplied by Bethyl, used in various techniques. Bioz Stars score: 93/100, based on 32 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/rabbit+polyclonal+anti+cpsf6/CPSF68+Antibody/bio_rxiv__2025__10__21__683728-227-21-24
Average 93 stars, based on 32 article reviews

rabbit polyclonal anti cpsf6 - by Bioz Stars, 2026-09

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Article Title: A twin UGUA motif directs the balance between gene isoforms through CFIm and the mTORC1 signaling pathway
Article Snippet: Reverse transcription and quantitative PCR (RT-qPCR) qPCR was performed using PerfeCTa SYBR Green SuperMix (QuantaBio) in triplicates.

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Article Title: Loss of CFIm activates YAP/TAZ and connects mRNA cleavage and polyadenylation inhibition to BRCAness
Article Snippet: 30∼60 μg lysates from culture cells were separated on 10% Bis-Tris Novex NuPAGE gels (Invitrogen) and transferred to nitrocellulose membrane following standard procedures. .. The following antibodies are used for western blotting: mouse monoclonal anti-alpha tubulin (Millipore, CP06), mouse monoclonal anti-HA (clone 16B12, BioLegend, 901501), rabbit polyclonal anti-CPSF6 (Bethyl Labs, A301-356A), YAP/TAZ (D24E4) Rabbit mAb (Cell Signaling Technology, 8418), YAP (D8H1X) Rabbit mAb (Cell Signaling Technology, 14074), Phospho-YAP (Ser127) (D9W2I) Rabbit mAb (Cell Signaling Technology, 13008), Phospho-Histone H2A.X (Ser139) (20E3) Rabbity mAb (Cell Signaling Technology, 9718), mouse monoclonal anti-NUDT21 (Proteintech, 66335-1-Ig), rabbit polyclonal anti-PD-L1/CD274 (Proteintech, 17952-1-ap), rabbit polyclonal anti-NEDD4 (Proteintech, 21698-1-ap), rabbit polyclonal anti-NEDD4L (Proteintech, 13690-1-ap), LATS1 (C66B5) Rabbit mAb (Cell Signaling Technology, 3477), LATS2 (D83D6) Rabbit mAb (Cell Signaling Technology, 5888). .. Quantitative PCR was performed using PerfeCTa SYBR Green SuperMix or PerfeCTa SYBR Green FastMix (QuantaBio) in triplicates.

Article Title: A twin UGUA motif directs the balance between gene isoforms through CFIm and the mTORC1 signaling pathway
Article Snippet: 20–60 μg lysates from culture cells or mouse tissues were separated on 10% Bis-Tris or 3~8% Tris-Acetate Novex NuPAGE gels (Invitrogen) and transferred to nitrocellulose membrane following standard procedures. .. The following antibodies are used for western blotting: mouse monoclonal anti-beta actin (ProteinTech, 66009-1-Ig), mouse monoclonal anti-alpha tubulin (Millipore, CP06), mouse monoclonal anti-HA (clone 16B12, BioLegend, 901501), rabbit polyclonal anti-TRIM9 (ProteinTech, 10786-1-AP), rabbit monoclonal anti-S6 ribosomal protein (Cell Signaling Technology, 2217), rabbit monoclonal anti-Phospho-S6 ribosomal protein (Ser240/244) (Cell Signaling Technology, 5364), rabbit polyclonal anti-CPSF6 (Bethyl Labs, A301-356A), rabbit polyclonal anti-BRD4 (Bethyl Labs, A301-985A), and rabbit monoclonal anti-BRD4 (Abcam, ab128874). .. qPCR was performed using PerfeCTa SYBR Green SuperMix (QuantaBio) in triplicates.

Article Title: A twin UGUA motif directs the balance between gene isoforms through CFIm and the mTORC1 signaling pathway
Article Snippet: 20~60μg lysates from culture cells or mouse tissues were separated on 10% Bis-Tris or 3~8% Tris-Acetate Novex NuPAGE gels (Invitrogen) and transferred to nitrocellulose membrane following standard procedures. .. The following antibodies are used for western blotting: mouse monoclonal anti-beta actin (Proteintech, 66009-1-Ig), mouse monoclonal anti-alpha tubulin (Millipore, CP06), mouse monoclonal anti-HA (clone 16B12, BioLegend, 901501), rabbit polyclonal anti-TRIM9 (Proteintech, 10786-1-AP), rabbit monoclonal anti-S6 ribosomal protein (Cell Signaling Technology, 2217), rabbit monoclonal anti-Phospho-S6 ribosomal protein (Ser240/244) (Cell Signaling Technology, 5364), rabbit polyclonal anti-CPSF6 (Bethyl Labs, A301-356A), rabbit polyclonal anti-BRD4 (Bethyl Labs, A301-985A), rabbit monoclonal anti-BRD4 (Abcam, ab128874). .. qPCR was performed using PerfeCTa SYBR Green SuperMix (QuantaBio) in triplicates.

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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.

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.

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.

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.

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.

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

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

Journal: iScience

Article Title: Multi-omics approach reveals posttranscriptionally regulated genes are essential for human pluripotent stem cells

doi: 10.1016/j.isci.2022.104289

Figure Lengend Snippet:

Article Snippet: Rabbit polyclonal anti-CPSF6 (1:100) , Proteintech , Cat# 15489-1-AP RRID: AB_10694140.

Techniques: Recombinant, Mass Spectrometry, Sequencing, Modification, Protease Inhibitor, Multiplex sample analysis, Multiplex Assay, Fractionation, Cell Culture, Bicinchoninic Acid Protein Assay, Lysis, Cloning, Gene Expression, SYBR Green Assay, Transfection, Ligation, Microarray, TaqMan Assay, Software, Cell Analysis

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