Journal: Toxicology and applied pharmacology

Article Title: Monoubiquitinated γ-H2AX: abundant product and specific biomarker for non-apoptotic DNA double-strand breaks

doi: 10.1016/j.taap.2018.07.007

Figure Lengend Snippet: Panels A-C used antibodies #1. (A) Impact of gel polyacrylamide percentage and transfer conditions to PVDF membranes. Treatments of H460 cells: 6 h with 3 mM hydroxyurea (HU), 6 or 16 h with 5 μM aphidicolin (Aphi) or 1 h with 5 μM bleomycin (Bleo). All γ-H2AX images had 5 s film exposures. (B) Ethanol effect in semi-dry transfer. IMR90 cells were treated with 40 μM etoposide (Eto) for 1 h. (C) Comparison of wet (pH 8.2 and pH 8.8) and semi-dry transfers using 12% ethanol-containing buffers (10 s film exposures for all γ-H2AX images). IMR90 cells were treated with 40 μM etoposide for 1 h. (D) Effectiveness of different antibodies using semi-dry transfer with 12% ethanol. Normal human keratinocytes were treated for 1 h with 30 μM etoposide or 0.5 μM bleomycin. Antibodies #1–3: 1:1000 dilutions and 5 s exposures, antibody #4: 1:1000 dilution and 60 s exposure. (E) Comparison of antibodies #1 and #5 (both at 1:1000 dilutions) in H460 cells treated with bleomycin for 1 h. (F) γ-H2AX blot (Ab#1) in control and H2AX-depleted IMR90 cells treated with 100 μM etoposide for 1h.

Article Snippet: IMR90, WI38, normal human neonatal keratinocytes (PCS-200–010), H460, A549, U2OS, mouse embryonic fibroblasts (MEFs, SCRC-1040), telomerase-immortalized human bronchial epithelial HBEC3 (CRL-4051), Daudi, HCT116, Ramos, Raji and CA46 were obtained from ATCC.

Techniques: Comparison, Control

Journal: bioRxiv

Article Title: KGF induces podosome formation via integrin-Erk1/2 signaling in human immortalized oral epithelial cells

doi: 10.1101/508416

Figure Lengend Snippet: KGF Mediated its Effects on HIOECs through Erk1/2 and Akt Signaling. (A and B) Western blot detection of the expression level of KGFR in HIOECs after KGF treatment, (C and D) the phosphorylation level of Jak2 and Stat3 after KGF stimulation for 15 min, and (E and F) the phosphorylation level of Erk1/2, p38, Jnk, and Akt after KGF stimulation for 15 min. n=3 independent experiments. Analysis of matrix protein expression in protein extracts from each group by ImageJ. (G) EdU labeling assay: the impact of U0126 and Ly294002 on the proliferation ability of HIOECs. (H) The quantification shown in the right graph indicates the mean ± SEM. (I) MTT assay: the impact of U0126 and Ly294002 on the proliferation ability of HIOECs. Data are expressed as the means ± SEM. * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: HIOECs at 3x10 4 cells/well were seeded in the 96-well plate and exposed to KGF (10 ng/mL; Proteintech) for 24 h. For MTT assays, the cells were incubated with 20 μL of MTT solution (5 mg/mL) at 37°C for 4 h. The supernatant was discarded, and 150 μL of dimethyl sulfoxide (DMSO) was added to each well.

Techniques: Western Blot, Expressing, Phospho-proteomics, Labeling, MTT Assay