Journal: Oncology Reports

Article Title: Pituitary tumor‑transforming gene 1 regulates the senescence and apoptosis of oral squamous cell carcinoma in a p21‑dependent DNA damage response manner

doi: 10.3892/or.2024.8794

Figure Lengend Snippet: PTTG1 expression in relation to cell proliferation, cell cycle and apoptosis in OSCC cells. (A) PTTG1 expression was analyzed by reverse transcription quantitative PCR in OSCC cells. (B) Protein expression of PTTG1 and PCNA in OSCC cell lines revealed by western blotting. The left panel shows the membranes stained with antibodies, with GAPDH as an internal control, and the right panel shows the relative quantification of protein expression. (C and D) Representative images of cell proliferation ability (left) and quantification (right) of PTTG1 using the EdU assay in the (C) HSC-2 and (D) SCC-9 cell lines (scale bars, 100 µm and 50 µm, respectively; original magnification, ×40). The percentages of HSC-2 and SCC-9 cells treated with VC or siR-PTTG1 were determined by EdU incorporation (green) and DAPI (blue). (E) Protein expression (left) and the fold change (right) of cell cycle markers, including cyclins D1, E, and B1 in OSCC cells. (F) Protein expression (left) and fold change (right) related to apoptosis markers, including Cas-7, c-Cas-7, and c-PARP in OSCC cells. *P<0.05, **P<0.01 and ***P<0.001 vs. VC using Student's t-test. All experiments were performed in triplicate. PTTG1, pituitary tumor-transforming gene 1; OSCC, oral squamous cell carcinoma; PCNA, proliferating cell nuclear antigen; VC, vehicle control; siR-PTTG1, small interfering RNA-PTTG1; EdU, 5-ethynyl-2′-deoxyuridine; Cas-7, Caspase-7; c-, cleaved-; PARP, poly (ADP-ribose) polymerase.

Article Snippet: Next, the membranes were incubated with the following primary antibodies overnight at 4°C: Rabbit anti-PTTG1 (cat. no. GTX111938; GeneTex, Inc.), anti-p21 (cat. no. 2947; Cell Signaling Technology, Inc.), anti-p53 (cat. no. 2527; Cell Signaling Technology, Inc.), anti-retinoblastoma (Rb; cat. no. 9390; Cell Signaling Technology, Inc.), anti-phosphorylated Rb (cat. no. 8180; Cell Signaling Technology, Inc.), anti-proliferating cell nuclear antigen (PCNA; cat. no. 2586; Cell Signaling Technology, Inc.), mouse anti-Caspase-7 (Cas-7; cat no. sc-56063; Santa Cruz Biotechnology, Inc.), anti-cleaved (c-) Cas-7 (cat. no. 8438; Cell Signaling Technology, Inc.), anti-c-poly (ADP-ribose) polymerase (PARP; cat. no. 5625; Cell Signaling Technology, Inc.), anti-PARP (cat. no. 9542; Cell Signaling Technology, Inc.), mouse anti-cyclin D1 (cat. no. sc-450; Santa Cruz Biotechnology, Inc.), mouse anti-cyclin E (cat. no. sc-247; Santa Cruz Biotechnology, Inc.), mouse anti-cyclin B1 (cat. no. sc-245; Santa Cruz Biotechnology, Inc.), anti-phosphorylated histone H2AX (γH2AX; cat. no. 80312; Cell Signaling Technology, Inc.), anti-phosphorylated ATR (cat. no. 2853; Cell Signaling Technology, Inc.) anti-ATR (cat. no. 2790; Cell Signaling Technology, Inc.), anti-phosphorylated ataxia telangiectasia mutant (p-ATM; cat. no. 13050; Cell Signaling Technology, Inc.) and anti-ATM (cat. no. 2873; Cell Signaling Technology, Inc.); all primary antibodies were diluted at 1:1,000 with BSA.

Techniques: Expressing, Reverse Transcription, Real-time Polymerase Chain Reaction, Western Blot, Staining, Control, EdU Assay, Small Interfering RNA

Journal: PLOS ONE

Article Title: Alpha-boswellic acid accelerates acute wound healing via NF-κB signaling pathway

doi: 10.1371/journal.pone.0308028

Figure Lengend Snippet: (A) Changes in TNFα protein level during the wound healing process at different concentrations of α-BA. The optimal concentration of α-BA was 100ug/ml. (B) Changes in IL-6 protein level during the wound healing process at different concentrations of α-BA. (C) Transwell migration assay to assess the impact of α-BA on the cell invasion capacity (48h). (D) Statistical graphs of Transwell migration assay results. (E) CCK8 assays to assess the impact of α-BA on the proliferative activity (48h). (F) Changes in Cyclin D1 mRNA levels induced by α-BA. (G) Representative images of cell cycle experiments. (H) Statistical graphs of cell cycle experiments. *P < 0.05 (control vs. LPS). ** P < 0.01 (control vs. LPS). *** P < 0.001 (control vs. LPS). # P < 0.05 (control vs. LPS + α-BA). ## P < 0.01 (control vs. LPS + α-BA). ### P < 0.001 (control vs. LPS + α-BA). ns: not statistically significant.

Article Snippet: The antibodies used included cyclin D1(1:1,000; Catalog no. ab16663; Abcam), p65 (1:1,000; Catalog no. ab16502; Abcam), IκBα (1:500; Catalog no. ab76429; Abcam), p-IκBα (1:10,000; Catalog no. ab133462; Abcam), GAPDH (1:5,000; Catalog no. ab8245; Abcam), and IgG secondary antibody (1:5,000; Catalog no. ab6789; Abcam).

Techniques: Concentration Assay, Transwell Migration Assay, Activity Assay, Control

Journal: bioRxiv

Article Title: The pro-apoptotic effect of chronic contractile activity-induced extracellular vesicles on Lewis Lung Carcinoma cells

doi: 10.1101/2024.08.29.610232

Figure Lengend Snippet: CCA-EVs were pretreated with Triton X-100 with or without proteinase K, and then co-cultured with LLC cells for 4 days. LLC cells were also treated with CON-EV, CCA-EV and PBS for 4 days, after which the level of senescence was measured. (A) Representative images and quantification of SA-β-gal activity. Scale bar: 200 µm at 10X magnification, and (B) western blot analysis of senescence markers, HMGB1, p16, and cyclin D1 in treated LLC cells. Data were analyzed using a one-way ANOVA in panel A with multiple comparisons corrected using Holm- Šídák post hoc test, and by an unpaired Student’s t-test in panel B. Data are expressed as scatter plots with mean (n = 4-6). Exact p values for significant (p<0.05) or close to statistically significant results is shown. The p-values for non-significant data are not shown.

Article Snippet: The following primary antibodies were used: mouse anti-p53 (2524, Cell Signaling, 1:500), rabbit anti-Apoptosis Inducing Factor (AIF) (5318, Cell Signaling, 1:1000), rabbit anti-caspase-3 (9662, Cell Signaling, 1:1000), rabbit anti-cleaved caspase-3 (9661, Cell Signaling, 1:1000), rabbit anti-Bcl-2 (3498, Cell Signaling, 1:1000), rabbit anti-Bax (2772, Cell Signaling, 1:1000), rabbit anti-cytochrome c (AHP2302, Bio-Rad Laboratories, 1:250), rabbit anti-High mobility group box 1 (HMGB1) (6893, Cell Signaling, 1:2000), mouse anti-p16 (SAB4500072, Sigma-Aldrich, 1:500), rabbit anti-cyclin D1 (2922, Cell Signaling, 1:1000), and mouse OXPHOS cocktail (45-8099, Invitrogen, 1:500).

Techniques: Cell Culture, Activity Assay, Western Blot