Journal: Journal of Pediatric Hematology/Oncology

Article Title: Interaction of E2F3a and CASP8AP2 Regulates Histone Expression and Chemosensitivity of Leukemic Cells

doi: 10.1097/MPH.0000000000002558

Figure Lengend Snippet: E2F3a and CASP8AP2 collaboratively affected H2A and H2B expression. A, Overexpression of E2F3a increased the expression of H2A and H2B in HEK-293T cells. B, E2F3a knockdown decreased the expression of H2A and H2B in HEK-293T cells. C, Overexpression of CASP8AP2 enhanced the expression of H2A and H2B. D, Downregulation of CASP8AP2 reduced the expression of histones in HEK-293T cells. E, The reduction in histones induced by the E2F3a knockdown could be reversed by the overexpression of CASP8AP2.

Article Snippet: A Western blot was carried out with the following antibodies: histone H2A and H2B antibodies (CST, USA), GAPDH antibody (SAB, USA), anti-FLAG antibody (Sigma, USA), and anti-GST antibody (CST).

Techniques: Expressing, Over Expression

Journal: Cell Death & Disease

Article Title: PVT1 inhibition stimulates anti-tumor immunity, prevents metastasis, and depletes cancer stem cells in squamous cell carcinoma

doi: 10.1038/s41419-023-05710-6

Figure Lengend Snippet: A Immunofluorescent staining and quantification of pH2A.X (red) in 4NQO-induced HNSCC treated with ASO PVT1. DAPI stained the nuclei blue. The white dotted line indicates the boundary between interstitial tissue and the tumor. Scale bar, 25 μm. Data are shown as the mean ± SD. n = 16. ** p < 0.01 using an unpaired Student’s t test. B Western blot of pH2A.X in HN6 and SCC15 cells with PVT1 KD. C , D Images and quantification of DNA Comet assays in HN6 and SCC15 cells treated with shPVT1 (>10 cells per group). Data are shown as the mean ± SD. Scale bar, 100 μm. ** p < 0.01 using an unpaired Student’s t test. E qRT-PCR analysis of IFNβ mRNA expression in HN6 and SCC15 cells with PVT1 KD. Data are shown as the mean ± SD. ** p < 0.01 using an unpaired Student’s t test. F qRT-PCR assessment of the expression levels of CXCL9, CXCL10, and CXCL11 in HN6 and SCC15 cells with PVT1 KD. Data are shown as the mean ± SD. ** p < 0.01 using an unpaired Student’s t-test.

Article Snippet: Primary antibodies recognizing the following proteins were used: CD24 (1:1000, Proteintech, Wuhan, China, Cat#18330-1-AP), SOX2 (1:1000, Cell Signaling Technology, Cat#14962), ALDH1 (1:1000, Cell Signaling Technology, Cat#54135), OCT4 (1:1000, Wanlei, Shenyang, China, WL02020), BMI1 (1:1000, Cell Signaling Technology, Cat#6964), YAP1 (1:1000, Cell Signaling Technology, Cat#14074), GAPDH (encoding glyceraldehyde-3-phosphate dehydrogenase) (1:20000, Proteintech, Cat#60004-1). anti-phospho Histone H2A.X (Ser139) (1:1000, Cell Signaling Technology, Cat#9718).

Techniques: Staining, Western Blot, Quantitative RT-PCR, Expressing

Journal: Scientific Reports

Article Title: Standard of care drugs do not modulate activity of senescent primary human lung fibroblasts

doi: 10.1038/s41598-023-30844-0

Figure Lengend Snippet: Lung fibroblasts isolated from normal and IPF patients exhibited a senescent phenotype after underwent through replicative senescence. Schematic presentation of normal and IPF lung fibroblasts transition from early to late passages ( a ). mRNA expression of senescence associated genes CDKN1A , CDKN1B , CDKN2A , and WNT16 in proliferating and senescent lung fibroblasts from normal or IPF patients ( b ). Representative images of SA-β-gal and the DNA damage marker γ-H2Ax co-staining is pronounced in senescent lung fibroblasts from normal or IPF patients ( c ). The scale bars indicate 100 μM (Spider-β-gal, γ-H2Ax, DAPI and merge) and 20 μM for amplified image. Quantification of SA-β-gal and γ-H2Ax total fluorescence/number of cells from proliferating and senescent lung fibroblasts from normal or IPF patients ( d ). SA-β-galactosidase staining is detected in senescent lung fibroblasts from normal and IPF patients but not in proliferating lung fibroblasts ( e ). Data are presented as mean ± SD (n = 3 or 5 per group). *p < 0.05, **p < 0.01 and ***p < 0.001 as indicated by the bars.

Article Snippet: After three washes, cells were permeabilized with 0.1% Triton X-100/PBS for 30 min and blocked in 1% BSA/PBS for 1 h. An anti- γ- H2A.x antibody (Cell Signaling Cat#2577L) diluted in 1% BSA/PBS was added to the cells and incubated overnight at 4 °C.

Techniques: Isolation, Expressing, Marker, Staining, Amplification, Fluorescence