Journal: Cell reports

Article Title: Non-canonical isoforms of the mRNA polyadenylation factor WDR33 regulate STING-mediated immune responses

doi: 10.1016/j.celrep.2024.113886

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Mouse anti-CPSF2 , Santa Cruz Biotechnology , Cat# sc-165983; RRID: AB_2084371.

Techniques: Virus, Recombinant, Negative Control, Software

Journal: Nucleic Acids Research

Article Title: THOC5 controls 3′end-processing of immediate early genes via interaction with polyadenylation specific factor 100 (CPSF100)

doi: 10.1093/nar/gku911

Figure Lengend Snippet: Upon stimulation with serum CPSF100 forms a complex with THOC5. ( A and B ) ER T2 Cre THOC5 (flox/flox) MEF cells were serum starved for 24 h and then stimulated with or without serum for 1 h. Nuclear fractions were isolated using RNase and then precipitated with antibody against THOC5 (A), or CPSF100 (B). Immunoprecipitates were analyzed by THOC5, CPSF100 or CFIm68-specific immunoblot. ( C ) Mouse NIH3T3 cells were transfected with CPSF100-specific siRNA and control siRNA and cell lysates were used for THOC5 and CSPF100-specific immunoblot. Actin was used as a loading control. ( D ) Sister cultures from (C) were incubated without serum for 24 h, and then cells were stimulated with serum for 30, 60 or 120 min. RNAs were isolated from each sample and semi-quantitative RT-PCR was performed using Id1, Id3, Ier2 , Egr1 and Gapdh -specific primers (Table ). Three independent experiments were performed and an example of representative data is shown here.

Article Snippet: Cells were treated with Tamoxifen (10 μM) for 24 h and then further incubated in growth medium for 24 h. After serum starvation for further 24 h, cells were stimulated with 20% serum for various times as indicated. siRNA against CPSF100 (SC-142546) and control siRNA (SC-37007) were from Santa Cruz Biotechnology Inc. (Santa Cruz, CA, USA).

Techniques: Isolation, Western Blot, Transfection, Control, Incubation, Quantitative RT-PCR

Journal: Nucleic Acids Research

Article Title: THOC5 controls 3′end-processing of immediate early genes via interaction with polyadenylation specific factor 100 (CPSF100)

doi: 10.1093/nar/gku911

Figure Lengend Snippet: THOC5 is required for recruiting of the CPSF100 to 3′end of THOC5 target gene. pCTAP (cTAP) and pCTAP carrying THOC5 cDNA (TAP-THOC5) were transfected into mouse NIH3T3 cells (Bait: THOC5 ( A )), or ER T2 THOC5 (flox/flox) MEF cells were treated with or without tamoxifen for 2 days (Baits: CPSF100 ( B ), RNApolymerase II ( C ), or CFIm68 ( D )). The cells were then incubated for 24 h in the presence of 20% FCS. After serum starvation for 24 h, cells were stimulated with (+) or without (−) serum for 1 h. After cross-linking by adding formaldehyde, protein and DNA were extracted and the chromatin was sheared by sonication. Cell extracts and binding fractions with streptavidin Sepharose or immunoprecipitates using CPSF100, RNApolymerase II or CFIm68 antibodies or control IgG were analyzed by Id3 (promoter region (-314- -199)and 3′UTR in exon 3 (1433-1571)) and Ier2 (promoter region (-701- -535) and 3′UTR region (1349-1522))-specific PCR (Table ; ChIP). The promoter region of each gene was described by Zhao et al . . Numbers represent nucleotide numbers from the initiation site for each gene. Data represent% input of each PCR reaction. Three independent experiments were performed.

Article Snippet: Cells were treated with Tamoxifen (10 μM) for 24 h and then further incubated in growth medium for 24 h. After serum starvation for further 24 h, cells were stimulated with 20% serum for various times as indicated. siRNA against CPSF100 (SC-142546) and control siRNA (SC-37007) were from Santa Cruz Biotechnology Inc. (Santa Cruz, CA, USA).

Techniques: Transfection, Incubation, Sonication, Binding Assay, Control

Journal: BMC Medicine

Article Title: Serum anti-DIDO1, anti-CPSF2, and anti-FOXJ2 antibodies as predictive risk markers for acute ischemic stroke

doi: 10.1186/s12916-021-02001-9

Figure Lengend Snippet: List of antibody biomarkers for atherosclerosis

Article Snippet: GST–FOXJ2 and GST–CPSF2 were purchased from Abnova (Taipei, Taiwan).

Techniques: Protein Array, Membrane, Binding Assay, Activation Assay

Journal: BMC Medicine

Article Title: Serum anti-DIDO1, anti-CPSF2, and anti-FOXJ2 antibodies as predictive risk markers for acute ischemic stroke

doi: 10.1186/s12916-021-02001-9

Figure Lengend Snippet: Presence of antibodies against DIDO1, FOXJ2, and CPSF2 in sera from a healthy donor (an HD) or a patient with transient ischemic attack (TIA) or acute ischemic stroke (AIS). Purified proteins of glutathione S-transferase (GST) (lane 1), GST–DIDO1 (1-275) (lane 2), GST–DIDO1 (271-545) (lane 3), GST–CPSF2 (lane 4), and GST–FOXJ2 (lane 5) were separated through sodium dodecyl sulfate-polyacrylamide gel electrophoresis, followed by western blotting analysis using anti-GST ( b ), anti-DIDO1 ( c ), anti-FOXJ2 ( d ), anti-CPSF2 ( e ), and sera from HD-#30017 ( f ), patients with AIS-#07684 ( g ), AIS-#07581 ( i ), and AIS-#07115 ( k ), and those with TIA-#07207 ( h ), TIA-#07175 ( j ), and TIA-#07060 ( l ). The Coomassie brilliant blue (CBB) staining profile is also shown in a . M, molecular weight marker

Article Snippet: GST–FOXJ2 and GST–CPSF2 were purchased from Abnova (Taipei, Taiwan).

Techniques: Purification, Polyacrylamide Gel Electrophoresis, Western Blot, Staining, Molecular Weight, Marker

Journal: BMC Medicine

Article Title: Serum anti-DIDO1, anti-CPSF2, and anti-FOXJ2 antibodies as predictive risk markers for acute ischemic stroke

doi: 10.1186/s12916-021-02001-9

Figure Lengend Snippet: Comparison of serum DIDO1-, CPSF2-, and FOXJ2-Ab levels between HDs and patients with TIA or AIS. GST-DIDO1:1-275 ( a ), biotinylated FOXJ2 peptide (bFOXJ2-426) ( d ), and biotinylated CPSF2 peptide (bCPSF2-607) ( g ) were used as the antigens. The AlphaLISA-determined serum antibody levels after subtraction of the levels against those of control GST are shown as box-whisker plots displaying the 10th, 20th, 50th, 80th, and 90th percentiles. P values were calculated by the Kruskal–Wallis test. * P < 0.05, ** P < 0.01, *** P < 0.001. The serum numbers of HDs, TIA, and AIS were 285, 92, and 464, respectively. The other information including total (male/female) numbers, average values, SDs, cutoff values, positive numbers, positive rates (%), and P values is summarized and shown in Table . Receiver operating characteristic curve (ROC) analysis was performed to assess the ability of DIDO1-Abs ( b , c ), FOXJ2-Abs ( e , f ), and CPSF2-Abs ( h , i ) to detect TIA and AIS. The numbers in the figures indicate the cutoff values for marker levels, and the numbers in parentheses indicate the sensitivity (left) and specificity (right). The areas under the curve (AUC), and 95% confidence intervals (CIs) are also shown in Table

Article Snippet: GST–FOXJ2 and GST–CPSF2 were purchased from Abnova (Taipei, Taiwan).

Techniques: Comparison, Control, Whisker Assay, Marker

Journal: BMC Medicine

Article Title: Serum anti-DIDO1, anti-CPSF2, and anti-FOXJ2 antibodies as predictive risk markers for acute ischemic stroke

doi: 10.1186/s12916-021-02001-9

Figure Lengend Snippet: Comparison of serum DIDO1-, FOXJ2-, and CPSF2-Ab levels between HDs and patients with acute myocardial infarction (AMI) or diabetes mellitus (DM) examined by AlphaLISA

Article Snippet: GST–FOXJ2 and GST–CPSF2 were purchased from Abnova (Taipei, Taiwan).

Techniques: Comparison

Journal: BMC Medicine

Article Title: Serum anti-DIDO1, anti-CPSF2, and anti-FOXJ2 antibodies as predictive risk markers for acute ischemic stroke

doi: 10.1186/s12916-021-02001-9

Figure Lengend Snippet: Receiver operating characteristic (ROC) analysis

Article Snippet: GST–FOXJ2 and GST–CPSF2 were purchased from Abnova (Taipei, Taiwan).

Techniques:

Journal: BMC Medicine

Article Title: Serum anti-DIDO1, anti-CPSF2, and anti-FOXJ2 antibodies as predictive risk markers for acute ischemic stroke

doi: 10.1186/s12916-021-02001-9

Figure Lengend Snippet: Comparison of serum DIDO1-Abs, FOXJ2-Abs, and CPSF2-Abs levels between HDs and patients with acute myocardial infarction (AMI) and diabetes mellitus (DM). GST-DIDO1:1-275 ( a ), bFOXJ2-426 ( d ), and bCPSF2-607 ( g ) were used as antigens. Serum antibody levels in HDs and patients with AMI and type 2 DM were determined using AlphaLISA and are shown as box-whisker plots, as described in the legend of Fig. . The same results are summarized in Table . Responses to DIDO1-Abs ( b , c ), FOXJ2-Abs ( e , f ), and CPSF2-Abs ( h , i ) were also evaluated using ROC analysis, and summarized in Table

Article Snippet: GST–FOXJ2 and GST–CPSF2 were purchased from Abnova (Taipei, Taiwan).

Techniques: Comparison, Whisker Assay

Journal: BMC Medicine

Article Title: Serum anti-DIDO1, anti-CPSF2, and anti-FOXJ2 antibodies as predictive risk markers for acute ischemic stroke

doi: 10.1186/s12916-021-02001-9

Figure Lengend Snippet: Comparison of the serum antibody levels of HDs versus those of patients with transient ischemic attack (TIA) or acute ischemic stroke (AIS)

Article Snippet: GST–FOXJ2 and GST–CPSF2 were purchased from Abnova (Taipei, Taiwan).

Techniques: Comparison

Journal: BMC Medicine

Article Title: Serum anti-DIDO1, anti-CPSF2, and anti-FOXJ2 antibodies as predictive risk markers for acute ischemic stroke

doi: 10.1186/s12916-021-02001-9

Figure Lengend Snippet: Comparison of serum antibody levels of HDs versus those of patients with chronic kidney disease (CKD)

Article Snippet: GST–FOXJ2 and GST–CPSF2 were purchased from Abnova (Taipei, Taiwan).

Techniques: Comparison

Journal: BMC Medicine

Article Title: Serum anti-DIDO1, anti-CPSF2, and anti-FOXJ2 antibodies as predictive risk markers for acute ischemic stroke

doi: 10.1186/s12916-021-02001-9

Figure Lengend Snippet: Comparison of serum DIDO1-Abs, FOXJ2-Abs, and CPSF2-Abs levels between HDs and patients with chronic kidney disease (CKD). Serum antibody levels against GST-DIDO1:1-275 protein ( a ), bFOXJ2-426 peptide ( e ), and bCPSF2-607 peptide ( i ) were compared between HDs and patients with CKD types 1, 2, and 3. The P values of CKD types 1, 2, and 3 versus HD controls are shown. Results are presented as described in the legend of Fig. . P values versus HD specimens are shown. The details are shown in Table . Responses to DIDO1-Abs ( b – d ), FOXJ2-Abs ( f – h ), and CPSF2-Abs ( j – l ) were also evaluated using the ROC analysis and are summarized in Table

Article Snippet: GST–FOXJ2 and GST–CPSF2 were purchased from Abnova (Taipei, Taiwan).

Techniques: Comparison

Journal: BMC Medicine

Article Title: Serum anti-DIDO1, anti-CPSF2, and anti-FOXJ2 antibodies as predictive risk markers for acute ischemic stroke

doi: 10.1186/s12916-021-02001-9

Figure Lengend Snippet: Correlation analysis of antibody levels against synthetic bDIDO1, bCPSF2, and bFOXJ2 peptides with data of subjects in the Sawara Hospital cohort

Article Snippet: GST–FOXJ2 and GST–CPSF2 were purchased from Abnova (Taipei, Taiwan).

Techniques:

Journal: BMC Medicine

Article Title: Serum anti-DIDO1, anti-CPSF2, and anti-FOXJ2 antibodies as predictive risk markers for acute ischemic stroke

doi: 10.1186/s12916-021-02001-9

Figure Lengend Snippet: Immunohistochemical staining of antigenic marker proteins in the atherosclerotic lesions. Surgically resected carotid atherosclerotic plaques were stained using immunohistochemistry. The antibodies used were anti-DIDO1 (Aviva Systems Biology), anti-FOXJ2 (Thermo Fisher Scientific), anti-CPSF2 (GeneTex), and anti-DHPS (Proteintech) antibodies for comparison. The tissue was also stained with antibodies against smooth muscle cell marker, vimentin (VIM) and smooth muscle actin (SMA), vascular endothelial cell marker, CD31 and CD34, and macrophage marker, CD68

Article Snippet: GST–FOXJ2 and GST–CPSF2 were purchased from Abnova (Taipei, Taiwan).

Techniques: Immunohistochemical staining, Staining, Marker, Immunohistochemistry, Comparison

Journal: BMC Medicine

Article Title: Serum anti-DIDO1, anti-CPSF2, and anti-FOXJ2 antibodies as predictive risk markers for acute ischemic stroke

doi: 10.1186/s12916-021-02001-9

Figure Lengend Snippet: Results of the Japan Public Health Center (JPHC) cohort samples

Article Snippet: GST–FOXJ2 and GST–CPSF2 were purchased from Abnova (Taipei, Taiwan).

Techniques: