Journal: Frontiers in oncology

Article Title: Mitotic Errors Promote Genomic Instability and Leukemia in a Novel Mouse Model of Fanconi Anemia.

doi: 10.3389/fonc.2021.752933

Figure Lengend Snippet: FIGURE 1 | Fancc-/-; Mad2+/- mice suffer from premature death and abnormal hematopoiesis. (A) Experimental design. Mad2 heterozygosity in Fancc-/- background is hypothesized to exacerbate mitotic abnormalities but not DNA damage repair response. Normal total white blood count (B), hemoglobin (C), platelet count (D), percentage of Cd11b+ and Gr1+ myeloid cells (E, G) and B220+ and Cd3+ lymphoid cells (F, H) in the peripheral blood of healthy-appearing Fancc-/-; Mad2+/- mice compared to age/sex-matched wt, Mad2+/- and Fancc-/- mice at 2-3 months of age. Well-appearing 16-week old Fancc-/-; Mad2+/- mice had the same bone marrow cellularity (I) and were able to form the same number of colonies in methylcellulose-based colony forming assays supplemented with progenitor growth factors (see Methods) (J) as age/sex-matched wt, Fancc-/- and Mad2+/- controls. Statistical analysis was performed by one-way ANOVA with Dunnett’s multiple comparison correction. No statistically significant differences were found. (K) Decreased cancer-free survival in Fancc-/-; Mad2+/- mice compared to wt, Mad2+/- and Fancc-/- age/sex-matched controls. Kaplan-Meier curves with p values determined by log-rank Mantel-Cox tests at 6-month and 24-month time points (n≥17 mice per genotype) are shown. Squares denote death due to hematopoietic malignancies; circles represent deaths from solid tumors. (L) Bone marrow, liver, and spleen infiltrates in representative Fancc-/-; Mad2+/- mice compared to wt controls. Insert (lower right) shows nodular splenomegaly in a representative leukemic Fancc-/-; Mad2+/- mouse. (M) Histopathological evidence of findings consistent with leukemia in five representative Fancc-/-; Mad2+/- mice. (N) flow cytometry demonstrating increased populations of large myeloid cells in a representative moribund Fancc-/-; Mad2+/- mouse. (O) Large myeloid blast cells, characterized by increased nucleus-to-cytoplasm ratio, irregular nuclei, and open chromatin from the peripheral blood. *p < 0.05, **p < 0.01, ****p < 0.0001.

Article Snippet: The following antibodies were used: anti-c-kit Frontiers in Oncology | www.frontiersin.org 3 (C19, Santa Cruz, 1:50), anti-cd11b (Novus, 1:50), CD3 (Dako, IR503), B220 (Clone RA3-6B2, BD Pharmingen).

Techniques: Comparison, Cytometry

Journal: Cell Death & Disease

Article Title: Transglutaminase 2 associated with PI3K and PTEN in a membrane-bound signalosome platform blunts cell death

doi: 10.1038/s41419-023-05748-6

Figure Lengend Snippet: A Relative mRNA expression of CD11b at the indicated days by RT-qPCR normalised to GAPDH ( n = 3). B Flow cytometry analysis of cell surface expression of the differentiation marker CD11b/CD18 ( n = 3). C Relative mRNA expression of CD11c measured by RT-qPCR normalised to GAPDH ( n = 3). D Flow cytometry analysis of cell surface expression of differentiation marker CD11c/CD18 ( n = 3). Measurements were conducted in triplicate; values were validated by Flowing software 2.5.1. Statistical analysis was conducted by two-way ANOVA (Bonferroni post hoc test; * p < 0.05, ** p < 0.01 and *** p < 0.001, **** p < 0.0001).

Article Snippet: F4/80 − cells were sorted and labelled with CD11c-PE and CD11b-FITC antibodies (1:25; R&D Systems) and APC-conjugated Annexin-V (Biolegend) for 15 min at 4 °C in the dark, and then analysed with a BD FACSAria III flow cytometer.

Techniques: Expressing, Quantitative RT-PCR, Flow Cytometry, Marker, Software