Journal:

Article Title: Expression of prostanoid receptors in human ductus arteriosus

doi: 10.1038/sj.bjp.0705092

Figure Lengend Snippet: Primer design for PCR amplification of prostanoid receptors and β-actin

Article Snippet: Abundance of receptor mRNA was expressed relative to β-actin mRNA. table ft1 table-wrap mode="anchored" t5 caption a7 Primer design for PCR amplification of prostanoid receptors and β -actin Open in a separate window

Techniques: Amplification

Journal:

Article Title: Expression of prostanoid receptors in human ductus arteriosus

doi: 10.1038/sj.bjp.0705092

Figure Lengend Snippet: Expression of prostanoid receptor mRNA in human DA. RNA was isolated from excised tissue, reverse-transcribed by oligo(dT) and PCR amplification for all tissue samples was repeated three times. A fragment of β-actin was amplified as internal control. A representative expression pattern of one tissue sample is shown, base pair markers (bp) are indicated on both sides.

Article Snippet: Abundance of receptor mRNA was expressed relative to β-actin mRNA. table ft1 table-wrap mode="anchored" t5 caption a7 Primer design for PCR amplification of prostanoid receptors and β -actin Open in a separate window

Techniques: Expressing, Isolation, Reverse Transcription, Amplification, Control

Journal:

Article Title: Expression of prostanoid receptors in human ductus arteriosus

doi: 10.1038/sj.bjp.0705092

Figure Lengend Snippet: Expression of the different prostanoid receptors relative to the expression of β-actin. RNA was isolated from excised tissue, reverse-transcribed by oligo(dT) and PCR amplification was performed for the indicated prostanoid receptors. Intensity of DNA bands is presented as percentage of amplification of β-actin fragment.

Article Snippet: Abundance of receptor mRNA was expressed relative to β-actin mRNA. table ft1 table-wrap mode="anchored" t5 caption a7 Primer design for PCR amplification of prostanoid receptors and β -actin Open in a separate window

Techniques: Expressing, Isolation, Reverse Transcription, Amplification

Journal: Blood Advances

Article Title: Dual ASXL1 and CSF3R mutations drive myeloid-biased stem cell expansion and enhance neutrophil differentiation

doi: 10.1182/bloodadvances.2024014362

Figure Lengend Snippet: Validation of a new mouse model of Csf3r and Asxl1 comutation. (A) Schematic of the recombination of the Csf3r -mutated allele. (B) Breeding scheme for the establishment of the Csf3r/Asxl1 -mutated cohort with appropriate controls producing all following 4 groups: control, Asxl1 Y588X , Csf3r T621I , and double mutant. (C) Polymerase chain reaction (PCR) validation of recombination of the Csf3r T621I allele. The recombined allele is 1024 bp long; the wild type one is 947 bp. (D) Primer design for the PCR validation of Csf3r T621I allele recombination in panel C. (E) PCR validation of the Asxl1 Y588X allele insertion. The primers used are P1 and P2 from Yang et al 18 giving a 350-bp amplicon for the wild-type allele and a 570-bp amplicon for the mutated allele. (F) Western blot revealing enhanced STAT3 and STAT5 phosphorylation in the bone marrow of mice with the Csf3r T621I allele.

Article Snippet: The recombined allele is 1024 bp long; the wild type one is 947 bp. (D) Primer design for the PCR validation of Csf3r T621I allele recombination in panel C. (E) PCR validation of the Asxl1 Y588X allele insertion.

Techniques: Biomarker Discovery, Control, Mutagenesis, Polymerase Chain Reaction, Amplification, Western Blot, Phospho-proteomics

Journal: Blood Advances

Article Title: Dual ASXL1 and CSF3R mutations drive myeloid-biased stem cell expansion and enhance neutrophil differentiation

doi: 10.1182/bloodadvances.2024014362

Figure Lengend Snippet: MYC overexpression reduces myeloid differentiation of Asxl1- and CSF3R -mutant cells. (A) Heat maps representing the normalized, row-scaled counts of the top 50 most significantly differentially expressed genes ( P adj < .05) between the control and double-mutant cells from the gene subset, Hallmark MYC targets. (B) Quantitative PCR (qPCR) for Myc before and after estrogen withdrawal (mean ± SEM; n = 3 per group). Significance was evaluated with a 2-way ANOVA with Sidak multiple comparison test. (C) qPCR for Myc expression in transduced double-mutant cells with a Tet-on MYC plasmid (mean ± SEM; n = 3 per group). Significance was evaluated with a 2-way ANOVA with Sidak multiple comparison test. (D) Representative flow cytometry plots of CD11b and GR1 at 48 hours post estrogen withdrawal of the double-mutant cells with a Tet-on MYC plasmid. These cells were treated with or without doxycycline 24 hours before estrogen withdrawal. Quantification of CSF3R-mCherry/Asxl1-GFP–positive cells reveals significant repression of CD11b + /GR1 + (mean ± SEM; n = 3 per group). Significance was evaluated with a 2-way ANOVA with Sidak multiple comparison test. ∗ P < .05, ∗∗ P < .01, ∗∗∗ P < .001, ∗∗∗∗ P < .0001.

Article Snippet: The recombined allele is 1024 bp long; the wild type one is 947 bp. (D) Primer design for the PCR validation of Csf3r T621I allele recombination in panel C. (E) PCR validation of the Asxl1 Y588X allele insertion.

Techniques: Over Expression, Mutagenesis, Control, Real-time Polymerase Chain Reaction, Comparison, Expressing, Plasmid Preparation, Flow Cytometry