Journal: Cell Death Discovery

Article Title: Hypoxia induced exosomal Circ-ZNF609 promotes pre-metastatic niche formation and cancer progression via miR-150-5p/VEGFA and HuR/ZO-1 axes in esophageal squamous cell carcinoma

doi: 10.1038/s41420-024-01905-8

Figure Lengend Snippet: A Transmission electron micrographs of extracellular vesicles derived from ECA109 and KYSE410. B The expression level of CD9, TSG101 and HSP70 (exosome specific markers) in isolated extracellular vehicles. C The nanoparticle concentration and size distribution of the extracellular vehicles derived from ECA109 and KYSE410. D Internalization of exosomes from ECA109 and KYSE410 by HUVECs. E HUVECs were treated with exosomes isolated from hypoxic cultured- ESCC cells (Hypo-Exo) or normoxic cultured- ESCC cells (Norm-Exo) respectively. The traverse of rhodamine labeled dextran probes through HUVECs monolayers were quantified through evaluating absorbance at 590 nm. F The expression level of angiogenesis related protein including VEGFA and Angiotensin II. HUVECs were incubated with Hypo-Exo from KYSE410, Norm-Exo from KYSE410 or without exosomes (control group) and then their circular RNA expressions profile was analyzed. G Heat map of the top 20 circular RN As which experienced an up-regulation (left) or down-regulation (right) in both Norm-Exo group and Hypo-Exo group. H Validation of the potentially dys-regulated circular RNAs in HUVECs internalized with exosomes. I HUVECs were transfected with circular RNA overexpression plasmid and an in vitro permeability assay was applied to determine the barrier function of endothelial cells. J The expression level of Circ-ZNF609 in ESCC cells exposed to indicate time in hypoxia. Data are expressed as mean ± SD. (* p < 0.05; ** p < 0.01; *** p < 0.001).

Article Snippet: Primary antibodies included CD9 (Abcam, ab236630), TSG101(Abcam, ab30871), CD63 (Abcam, ab271286), Argonaute-2 (Abcam, ab186733), HuR (Abcam, ab200342), VEGFA (Abcam, ab46154), Angiotensin II (Novus), β-Actin (Abcam, ab8226), ZO-1 (CST, 5406), Occludin (CST, 91131) and Claudin 1(CST, 13255).

Techniques: Transmission Assay, Derivative Assay, Expressing, Isolation, Concentration Assay, Cell Culture, Labeling, Incubation, Control, Biomarker Discovery, Transfection, Over Expression, Plasmid Preparation, In Vitro, Permeability

Journal: NPG Asia Materials

Article Title: Spatiotemporal regulation of endogenous MSCs using a functional injectable hydrogel system for cartilage regeneration

doi: 10.1038/s41427-021-00339-3

Figure Lengend Snippet: Fig. 10 MSC homing in cartilage defects at 7 days after the operation. A Immunofluorescent staining for CD44 and CD90. B, C Quantitative results for CD44- and CD90-positive cells.

Article Snippet: At 1 week postoperatively, samples were assessed by immunofluorescence staining for CD44 (1:100, NBP222530F, Novus) and CD90 (1:100, ab225, Abcam) to identify endogenous MSC recruitment.

Techniques: Staining

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