Journal: iScience

Article Title: Chromodomain helicase DNA-binding domain 2 maintains spermatogonial self-renewal by promoting chromatin accessibility and mRNA stability

doi: 10.1016/j.isci.2022.105552

Figure Lengend Snippet:

Article Snippet: Rabbit polyclonal anti-CSTF1 , Proteintech , Cat# 10064-2-AP Lot:00000083 RRID: AB_2261145.

Techniques: Recombinant, Embryo Culture, CCK-8 Assay, Staining, TUNEL Assay, Apoptosis Assay, Mass Spectrometry, Plasmid Preparation, Software

Journal: iScience

Article Title: Targeting the mRNA endonuclease CPSF73 inhibits breast cancer cell migration, invasion, and self-renewal

doi: 10.1016/j.isci.2022.104804

Figure Lengend Snippet:

Article Snippet: Rabbit polyclonal anti-CstF50 , Bethyl , Cat#A301-250A; RRID: AB_890586.

Techniques: Recombinant, Cell Viability Assay, shRNA, Control, Plasmid Preparation, Software

Journal: Frontiers in Cardiovascular Medicine

Article Title: Utility of atherosclerosis-associated serum antibodies against colony-stimulating factor 2 in predicting the onset of acute ischemic stroke and prognosis of colorectal cancer

doi: 10.3389/fcvm.2023.1042272

Figure Lengend Snippet: Detection of serum antibodies against CSF2. Presence of antibodies against CSF2 in the serum samples of patients with acute ischemic stroke (AIS), diabetes mellitus (DM), esophageal cancer (EC), and colorectal cancer (CRC). Purified GST-CSF2 (lane 1) and GST (lane 2) proteins were separated on sodium dodecyl sulfate–polyacrylamide gels, followed by staining with Coomassie Brilliant Blue (CBB) or western blotting using anti-GST (αGST), anti-CSF2 (αCSF2), and serum samples from patients with AIS (07544, 07684), DM (22226), EC (EC-6), and CRC (Co-58) and from a healthy donor (HD)(09101). The full, non-adjusted image of is shown in the . Molecular weights are presented in the left lane. Arrows indicate the positions of GST-CSF2 (44.6 kDa) and GST (26 kDa). CSF2, colony-stimulating factor 2.

Article Snippet: The membranes were blocked with 0.1% dry milk in Tris-buffered saline [150 mM NaCl, 20 mM Tris–HCl (pH 7.6), and 0.1% Tween-20; TBS-T] and subsequently incubated with anti-GST antibodies (goat, ab6613, Abcam, Cambridgeshire, United Kingdom), anti-CSF2 antibodies (rabbit, sc-37,753, Santa Cruz Biotechnology, Dallas, TX), or HD and patient sera diluted at 1:1000.

Techniques: Purification, Staining, Western Blot

Journal: Frontiers in Cardiovascular Medicine

Article Title: Utility of atherosclerosis-associated serum antibodies against colony-stimulating factor 2 in predicting the onset of acute ischemic stroke and prognosis of colorectal cancer

doi: 10.3389/fcvm.2023.1042272

Figure Lengend Snippet: Comparison of the s-CSF2-Ab and s-CSF2pep-Ab levels in patients with AIS and TIA and in HDs. (A–B) Levels of serum antibodies against (A) s-CSF2-Ab and (B) s-CSF2pep-Ab determined using amplified luminescence proximity homogeneous assay-linked immunosorbent assay (AlphaLISA), calculated by subtracting the levels of antibodies against the control GST. Scatter dot plots of the s-CSF2-Ab and s-CSF2pep-Ab levels are also presented. Bars represent averages ± standard deviation (SD). ** P < 0.05; * P < 0.01; ns, not significant, Kruskal–Wallis test. (C–E) A receiver operating characteristic (ROC) curve analysis was conducted to assess the ability of s-CSF2-Ab to detect AIS (C) and that of s-CSF2pep-Ab to detect AIS (D) and TIA (E) . The numbers indicate the cutoff values for the indicated markers, and the numbers in parentheses indicate sensitivity (left) and specificity (right). The areas under the ROC curve (AUC) and 95% confidence intervals (CIs) are also shown. AIS, acute ischemic stroke; HD, healthy donor; CSF2-Ab, CSF2 antibody; CSF2pep-Ab, CSF2 peptide antibody; TIA, transient ischemic attack.

Article Snippet: The membranes were blocked with 0.1% dry milk in Tris-buffered saline [150 mM NaCl, 20 mM Tris–HCl (pH 7.6), and 0.1% Tween-20; TBS-T] and subsequently incubated with anti-GST antibodies (goat, ab6613, Abcam, Cambridgeshire, United Kingdom), anti-CSF2 antibodies (rabbit, sc-37,753, Santa Cruz Biotechnology, Dallas, TX), or HD and patient sera diluted at 1:1000.

Techniques: Comparison, Amplification, Control, Standard Deviation

Journal: Frontiers in Cardiovascular Medicine

Article Title: Utility of atherosclerosis-associated serum antibodies against colony-stimulating factor 2 in predicting the onset of acute ischemic stroke and prognosis of colorectal cancer

doi: 10.3389/fcvm.2023.1042272

Figure Lengend Snippet: Comparison of the s-CSF2-Ab and s-CSF2pep-Ab levels in HDs versus those in patients with AIS and TIA.

Article Snippet: The membranes were blocked with 0.1% dry milk in Tris-buffered saline [150 mM NaCl, 20 mM Tris–HCl (pH 7.6), and 0.1% Tween-20; TBS-T] and subsequently incubated with anti-GST antibodies (goat, ab6613, Abcam, Cambridgeshire, United Kingdom), anti-CSF2 antibodies (rabbit, sc-37,753, Santa Cruz Biotechnology, Dallas, TX), or HD and patient sera diluted at 1:1000.

Techniques: Comparison

Journal: Frontiers in Cardiovascular Medicine

Article Title: Utility of atherosclerosis-associated serum antibodies against colony-stimulating factor 2 in predicting the onset of acute ischemic stroke and prognosis of colorectal cancer

doi: 10.3389/fcvm.2023.1042272

Figure Lengend Snippet: Comparing the s-CSF2-Ab and s-CSF2pep-Ab levels between HDs and patients with AMI or DM.

Article Snippet: The membranes were blocked with 0.1% dry milk in Tris-buffered saline [150 mM NaCl, 20 mM Tris–HCl (pH 7.6), and 0.1% Tween-20; TBS-T] and subsequently incubated with anti-GST antibodies (goat, ab6613, Abcam, Cambridgeshire, United Kingdom), anti-CSF2 antibodies (rabbit, sc-37,753, Santa Cruz Biotechnology, Dallas, TX), or HD and patient sera diluted at 1:1000.

Techniques:

Journal: Frontiers in Cardiovascular Medicine

Article Title: Utility of atherosclerosis-associated serum antibodies against colony-stimulating factor 2 in predicting the onset of acute ischemic stroke and prognosis of colorectal cancer

doi: 10.3389/fcvm.2023.1042272

Figure Lengend Snippet: Comparison of the s-CSF2-Ab and s-CSF2pep-Ab levels between HDs and patients with AMI and DM. ( A,B ) The levels of s-CSF2-Ab and s-CSF2pep-Ab in HDs and patients with acute myocardial infarction (AMI) ( A ) and diabetes mellitus (DM) ( B ) were determined using AlphaLISA. The bars represent averages ± SD. *** P < 0.001, Mann–Whitney U test. ( D,E ) The ROC curves to assess the ability of s-CSF2-Ab to predict AMI and DM are presented in ( D ) and ( E ), respectively. ( F,G ) The ability of s-CSF2pep-Ab to predict AMI ( E ) and DM ( F ) was also evaluated via ROC analysis. Comparison of the overall survival in patients with DM between the s-CSF2-Ab positive (s-CSF2-Ab + ) and s-CSF2-Ab negative (s-CSF2-Ab − ) groups ( P = 0.019; C ) and between the s-CSF2pep-Ab positive (s-CSF2-Ab + ) and s-CSF2pep-Ab negative (s-CSF2-Ab − ) groups ( P = 0.034; C ). Statistical analyzes were performed by the Log-Rank test between two groups.

Article Snippet: The membranes were blocked with 0.1% dry milk in Tris-buffered saline [150 mM NaCl, 20 mM Tris–HCl (pH 7.6), and 0.1% Tween-20; TBS-T] and subsequently incubated with anti-GST antibodies (goat, ab6613, Abcam, Cambridgeshire, United Kingdom), anti-CSF2 antibodies (rabbit, sc-37,753, Santa Cruz Biotechnology, Dallas, TX), or HD and patient sera diluted at 1:1000.

Techniques: Comparison, MANN-WHITNEY

Journal: Frontiers in Cardiovascular Medicine

Article Title: Utility of atherosclerosis-associated serum antibodies against colony-stimulating factor 2 in predicting the onset of acute ischemic stroke and prognosis of colorectal cancer

doi: 10.3389/fcvm.2023.1042272

Figure Lengend Snippet: Association of the s-CSF2-Ab and s-CSF2pep-Ab levels with CKD. The levels of s-CSF-Ab ( A ) and s-CSF2pep-Ab ( B ) in HDs and in patients with diabetic CKD (CKD type 1), nephrosclerosis (CKD type 2), and glomerulonephritis (CKD type 3) were examined using AlphaLISA. The bars represent averages ± SD. * P < 0.05; ** P < 0.01; *** P < 0.001, Kruskal–Wallis test. The data are summarized in . ( C–E ) ROC analysis to determine the ability of s-CSF2-Ab to predict CKD type 1 ( C ), type 2 ( D ), and type 3 ( E ). ( F–H ) ROC analysis to determine the ability of s-CSF2pep-Ab to predict CKD type 1 ( F ), type 2 ( G ), and type 3 ( H ). The numbers in the graphs are the same as those presented . CKD, chronic kidney disease.

Article Snippet: The membranes were blocked with 0.1% dry milk in Tris-buffered saline [150 mM NaCl, 20 mM Tris–HCl (pH 7.6), and 0.1% Tween-20; TBS-T] and subsequently incubated with anti-GST antibodies (goat, ab6613, Abcam, Cambridgeshire, United Kingdom), anti-CSF2 antibodies (rabbit, sc-37,753, Santa Cruz Biotechnology, Dallas, TX), or HD and patient sera diluted at 1:1000.

Techniques:

Journal: Frontiers in Cardiovascular Medicine

Article Title: Utility of atherosclerosis-associated serum antibodies against colony-stimulating factor 2 in predicting the onset of acute ischemic stroke and prognosis of colorectal cancer

doi: 10.3389/fcvm.2023.1042272

Figure Lengend Snippet: The s-CSF2-Ab and s-CSF2pep levels associated with CKD.

Article Snippet: The membranes were blocked with 0.1% dry milk in Tris-buffered saline [150 mM NaCl, 20 mM Tris–HCl (pH 7.6), and 0.1% Tween-20; TBS-T] and subsequently incubated with anti-GST antibodies (goat, ab6613, Abcam, Cambridgeshire, United Kingdom), anti-CSF2 antibodies (rabbit, sc-37,753, Santa Cruz Biotechnology, Dallas, TX), or HD and patient sera diluted at 1:1000.

Techniques:

Journal: Frontiers in Cardiovascular Medicine

Article Title: Utility of atherosclerosis-associated serum antibodies against colony-stimulating factor 2 in predicting the onset of acute ischemic stroke and prognosis of colorectal cancer

doi: 10.3389/fcvm.2023.1042272

Figure Lengend Snippet: Association between s-CSF2-Ab levels and the data from participants in the Sawara Hospital cohort.

Article Snippet: The membranes were blocked with 0.1% dry milk in Tris-buffered saline [150 mM NaCl, 20 mM Tris–HCl (pH 7.6), and 0.1% Tween-20; TBS-T] and subsequently incubated with anti-GST antibodies (goat, ab6613, Abcam, Cambridgeshire, United Kingdom), anti-CSF2 antibodies (rabbit, sc-37,753, Santa Cruz Biotechnology, Dallas, TX), or HD and patient sera diluted at 1:1000.

Techniques:

Journal: Frontiers in Cardiovascular Medicine

Article Title: Utility of atherosclerosis-associated serum antibodies against colony-stimulating factor 2 in predicting the onset of acute ischemic stroke and prognosis of colorectal cancer

doi: 10.3389/fcvm.2023.1042272

Figure Lengend Snippet: Correlation analysis of s-CSF2-Ab levels with data from subjects in the Sawara Hospital cohort.

Article Snippet: The membranes were blocked with 0.1% dry milk in Tris-buffered saline [150 mM NaCl, 20 mM Tris–HCl (pH 7.6), and 0.1% Tween-20; TBS-T] and subsequently incubated with anti-GST antibodies (goat, ab6613, Abcam, Cambridgeshire, United Kingdom), anti-CSF2 antibodies (rabbit, sc-37,753, Santa Cruz Biotechnology, Dallas, TX), or HD and patient sera diluted at 1:1000.

Techniques:

Journal: Frontiers in Cardiovascular Medicine

Article Title: Utility of atherosclerosis-associated serum antibodies against colony-stimulating factor 2 in predicting the onset of acute ischemic stroke and prognosis of colorectal cancer

doi: 10.3389/fcvm.2023.1042272

Figure Lengend Snippet: Comparison of the s-CSF2-Ab levels between HDs and patients with cancers. ( A ) Scatter dot plots of the s-CSF2-Ab levels in HDs and in patients with EC, gastric cancer (GC), CRC, lung cancer (LC), and mammary cancer (MC). The results are presented as described in the legend of . ** P < 0.01; *** P < 0.001; ns, not significant. ROC analysis was conducted to assess the ability of s-CSF2-Abs to detect EC ( B ), GC ( C ), CRC ( D ), and LC ( E ).

Article Snippet: The membranes were blocked with 0.1% dry milk in Tris-buffered saline [150 mM NaCl, 20 mM Tris–HCl (pH 7.6), and 0.1% Tween-20; TBS-T] and subsequently incubated with anti-GST antibodies (goat, ab6613, Abcam, Cambridgeshire, United Kingdom), anti-CSF2 antibodies (rabbit, sc-37,753, Santa Cruz Biotechnology, Dallas, TX), or HD and patient sera diluted at 1:1000.

Techniques: Comparison

Journal: Frontiers in Cardiovascular Medicine

Article Title: Utility of atherosclerosis-associated serum antibodies against colony-stimulating factor 2 in predicting the onset of acute ischemic stroke and prognosis of colorectal cancer

doi: 10.3389/fcvm.2023.1042272

Figure Lengend Snippet: Association of s-CSF2-Ab levels with cancer development.

Article Snippet: The membranes were blocked with 0.1% dry milk in Tris-buffered saline [150 mM NaCl, 20 mM Tris–HCl (pH 7.6), and 0.1% Tween-20; TBS-T] and subsequently incubated with anti-GST antibodies (goat, ab6613, Abcam, Cambridgeshire, United Kingdom), anti-CSF2 antibodies (rabbit, sc-37,753, Santa Cruz Biotechnology, Dallas, TX), or HD and patient sera diluted at 1:1000.

Techniques:

Journal: Frontiers in Cardiovascular Medicine

Article Title: Utility of atherosclerosis-associated serum antibodies against colony-stimulating factor 2 in predicting the onset of acute ischemic stroke and prognosis of colorectal cancer

doi: 10.3389/fcvm.2023.1042272

Figure Lengend Snippet: Comparison of survival according to s-CSF2-Ab positivity in patients with CRC after surgery. ( A ) Comparison of the postoperative survival between the s-CSF2-Ab + and s-CSF2-Ab − patients with CRC ( P = 0.011). ( B ) Comparison of the overall survival between the p53-Ab + (≥1.3 U/ml) and p53-Ab − (<1.3 U/ml) patients with CRC ( P = 0.952). ( C ) Comparison of the overall survival between the s-CSF2-Ab + and s-CSF2-Ab − patients with CRC among those with p53-Ab ≥1.3 U/ml ( P = 0.463). ( D ) Comparison of the overall survival between the s-CSF2-Ab + and s-CSF2-Ab − patients with CRC among those with p53-Ab <1.3 U/ml ( P = 0.004). The cutoff values in patients with CRC were determined via the ROC analysis presented in . Statistical analyzes were performed by the Log-Rank test between two groups.

Article Snippet: The membranes were blocked with 0.1% dry milk in Tris-buffered saline [150 mM NaCl, 20 mM Tris–HCl (pH 7.6), and 0.1% Tween-20; TBS-T] and subsequently incubated with anti-GST antibodies (goat, ab6613, Abcam, Cambridgeshire, United Kingdom), anti-CSF2 antibodies (rabbit, sc-37,753, Santa Cruz Biotechnology, Dallas, TX), or HD and patient sera diluted at 1:1000.

Techniques: Comparison

Journal: Frontiers in Cardiovascular Medicine

Article Title: Utility of atherosclerosis-associated serum antibodies against colony-stimulating factor 2 in predicting the onset of acute ischemic stroke and prognosis of colorectal cancer

doi: 10.3389/fcvm.2023.1042272

Figure Lengend Snippet: Association between s-CSF2-Ab levels and data from participants in the Toho University Hospital cohort.

Article Snippet: The membranes were blocked with 0.1% dry milk in Tris-buffered saline [150 mM NaCl, 20 mM Tris–HCl (pH 7.6), and 0.1% Tween-20; TBS-T] and subsequently incubated with anti-GST antibodies (goat, ab6613, Abcam, Cambridgeshire, United Kingdom), anti-CSF2 antibodies (rabbit, sc-37,753, Santa Cruz Biotechnology, Dallas, TX), or HD and patient sera diluted at 1:1000.

Techniques:

Journal: Frontiers in Cardiovascular Medicine

Article Title: Utility of atherosclerosis-associated serum antibodies against colony-stimulating factor 2 in predicting the onset of acute ischemic stroke and prognosis of colorectal cancer

doi: 10.3389/fcvm.2023.1042272

Figure Lengend Snippet: Diagram of the relationship between s-CSF2-Ab and p53 in cancer and atherosclerosis-related diseases. Arrows represent the downstream activation, and blocked arrows represent inhibitory action. Created with BioRender.com . MDM2 , mouse double minute 2 homolog.

Article Snippet: The membranes were blocked with 0.1% dry milk in Tris-buffered saline [150 mM NaCl, 20 mM Tris–HCl (pH 7.6), and 0.1% Tween-20; TBS-T] and subsequently incubated with anti-GST antibodies (goat, ab6613, Abcam, Cambridgeshire, United Kingdom), anti-CSF2 antibodies (rabbit, sc-37,753, Santa Cruz Biotechnology, Dallas, TX), or HD and patient sera diluted at 1:1000.

Techniques: Activation Assay

Journal: bioRxiv

Article Title: Uncoupling from transcription protects polyadenylation site cleavage from inhibition by DNA damage

doi: 10.1101/2022.05.26.492040

Figure Lengend Snippet: (A) Western blot of pre-mRNA 3’-end processing factors in response to UV treatment (40 J/m 2 ) of A549 cells, followed by16 hours of recovery. GAPDH was used as a loading control. (B) Western blot analysis of PCF11 expression in A549 cells transfected for 48 hours with two different siRNA targeting PCF11 prior to exposure to UV. (C) RT-qPCR measuring relative PCF11 mRNA level in A549 cells in response to UV treatment (40 J/m 2 ). The expression was normalized to RP18S. * P <0.05; unpaired t-test. (D) Scheme representing the RT-qPCR strategy for assessing pre-mRNA 3′-end processing efficiency. Primers for uncleaved pre-mRNAs are located downstream of the polyadenylation site, while primers that detect both processed (cleaved) and unprocessed RNAs (uncleaved) amplify upstream of the polyadenylation site. The ratio of uncleaved/total (cleaved+uncleaved) indicates the processing efficiency, where a greater uncleaved/total ratio represents a reduced processing. (E) RT-qPCR assay on nuclear RNA for assessing the uncleaved/total ratio of p53 and TATA-binding protein ( TBP ) pre-mRNAs in A549 cells transfected for 48 hours with two different siRNA targeting PCF11 prior to exposure (UV+) or not (UV-) to UV irradiation (40 J/m 2 ). (F) RT-qPCR assay on nuclear RNA for assessing the uncleaved/total ratio of p53 pre-mRNA in A549 cells transfected for 48 hours with siRNAs targeting the Cstf64, CF1m25 and CPSF160 and exposed to UV irradiation (40 J/m 2 ).

Article Snippet: Primary antibodies used in this study from Bethyl Laboratories were: CPSF160, CPSF100, CPSF73, CPSF30, CstF77, CstF64, CstF50, CFIm68 and CFIm59.

Techniques: Western Blot, Control, Expressing, Transfection, Quantitative RT-PCR, Binding Assay, Irradiation