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: Cell Biology and Toxicology

Article Title: Antitumorigenic potential of Lactobacillus -derived extracellular vesicles: p53 succinylation and glycolytic reprogramming in intestinal epithelial cells via SIRT5 modulation

doi: 10.1007/s10565-024-09897-y

Figure Lengend Snippet: LEV-mediated regulation of the SIRT5/p53 axis affects proliferation and glycolysis in intestinal epithelial cells, influencing colon tumor formation. Note: ( A ) Schematic diagram illustrating the construction of a mouse model of colitis-associated tumors; ( B ) co-immunoprecipitation (co-IP) experiment detecting the acetylation level of p53 in mouse colonic polyp tissues from each group; ( C ) statistical analysis of the number and burden of colonic polyps in each group of mice; ( D ) immunohistochemical staining detecting the positive expression of Ki67 protein in mouse colonic polyp tissues from each group (scale bar = 100 μm); ( E ) immunohistochemical staining detecting the positive expression of cell cycle-related proteins Cyclin D1 and p27 in mouse colonic polyp tissues from each group (scale bar = 100 μm); ( F ) glucose uptake in mouse colonic polyp tissues from each group; ( G ) lactate production in mouse colonic mucosal tissues from each group; ( H ) Western blot detecting the expression of glycolytic rate-limiting enzymes GLUT1 and HKII in mouse colonic mucosal tissues from each group. * represents a difference compared to the WT group (P < 0.05), # represents a difference compared to the LEVs + WT group (P < 0.05), 6 mice per group

Article Snippet: The primary antibodies used were rabbit anti-Ki67 (1:2000, 28,074–1-AP, Proteintech, Wuhan, China), mouse anti-Cyclin D1 (1:500, 60,186–1-Ig, Proteintech, Wuhan, China), rabbit anti-p27 (1:200, 25,614–1-AP, Proteintech, Wuhan, China), and rabbit anti-SIRT5 (1:200, 15,122–1-AP, Proteintech, Wuhan, China).

Techniques: Immunoprecipitation, Co-Immunoprecipitation Assay, Immunohistochemical staining, Staining, Expressing, Western Blot

Journal: Journal of Xenobiotics

Article Title: Chronic Exposure to Both Electronic and Conventional Cigarettes Alters Ileum and Colon Turnover, Immune Function, and Barrier Integrity in Mice

doi: 10.3390/jox14030053

Figure Lengend Snippet: Effects of chronic electronic and conventional cigarette exposure on ileum- and colon-proliferation signalling pathways. Transcript levels of markers of major proliferation pathways in ileum ( A ) and colon ( B ). n = 14 per group. Transcript levels of cyclin D1 pathway genes in ileum ( C ) and colon ( D ). n = 14 per group. Cyclin D1 immunostaining, with higher magnification on the right, and quantification of cyclin D1 mean optical density in ileum ( E ) and colon ( F ). Cyclin D1 stain: brown. Nuclear stain: blue. n = 5 per group. * p < 0.05, ** p < 0.01, *** p < 0.005 and **** p < 0.001 compared to the control group (Air), as determined by the Mann–Whitney U test.

Article Snippet: Rabbit anti-mouse Ki-67 primary antibody (MA5-14520, Invitrogen; Fisher Scientific, Illkirch, France 1/100 dilution), rabbit anti-mouse cyclin D1 primary antibody (#55506, Cell Signaling Technology; Danvers, USA 1/400 dilution), rabbit anti-mouse ZO-1 primary antibody (#61-7300, Invitrogen; 1/200 dilution), and rabbit anti-mouse occludin primary antibody (#91131, Cell Signaling; 1/200 dilution) were incubated overnight at 4 °C.

Techniques: Immunostaining, Staining, Control, MANN-WHITNEY

Journal: The Journal of Veterinary Medical Science

Article Title: Activation of the Wnt/β-catenin signaling pathway and CTNNB1 mutations in canine intestinal epithelial proliferative lesions

doi: 10.1292/jvms.24-0125

Figure Lengend Snippet: (a–c) Colorectal inflammatory polyp (CIP), case 65. Immunohistochemistry. Inset: higher magnification. ( a ) Nuclear and cytoplasmic β-catenin immunolabelling in dysplastic epithelium. ( b ) Nuclear immunolabeling for c-Myc in dysplastic epithelium. ( c ) Nuclear immunolabeling for Cyclin D1 in dysplastic epithelium. (d–f) Intestinal epithelial tumor (IET), case 26. Immunohistochemistry. Inset: higher magnification. ( d ) Nuclear and cytoplasmic immunolabeling for β-catenin in tumor cells. ( e ) Nuclear immunolabeling for c-Myc in tumor cells. ( f ) Nuclear immunolabeling for Cyclin D1 in tumor cells. (g–l) Immunolabeling scores of c-Myc and Cyclin D1 and Ki-67 index in β-catenin (+) and in β-catenin (−) cases. (g–i) CIPs: ( g ) The c-Myc score was significantly higher in β-catenin (+) cases than in β-catenin (−) cases. ( h ) The Cyclin D1 score was significantly higher in β-catenin (+) cases than in β-catenin (−) cases. ( i ) The Ki-67 index was higher in β-catenin (+) cases than in β-catenin (−) cases. (j-l) IETs: ( j ) The c-Myc score was significantly higher in β-catenin (+) cases than in β-catenin (−) cases. ( k ) The Cyclin D1 score was significantly higher in β-catenin (+) cases than in β-catenin (−) cases. ( l ) The Ki-67 index was higher in β-catenin (+) cases than in β-catenin (−) cases.

Article Snippet: The following primary antibodies were used: mouse anti-β-catenin antibody (1:1,000, clone 14/beta-catenin; BD Transduction Laboratories, Franklin Lakes, NJ, USA), rabbit anti-c-Myc antibody (1:250, clone Y69; Abcam, Cambridge, UK), mouse anti-cyclin D1 antibody (1:50, clone DCS-6; Santa Cruz Biotechnology, Dallas, TX, USA), and mouse anti-Ki-67 antibody (ready to use, clone MIB-1; Dako, Tokyo, Japan).

Techniques: Immunohistochemistry, Immunolabeling

Journal: Nagoya Journal of Medical Science

Article Title: Protective effect of Sasa veitchii extract against all-trans-retinoic acid-induced inhibition of proliferation of cultured human palate cells

doi: 10.18999/nagjms.86.2.223

Figure Lengend Snippet: Involvement with apoptosis and cell cycle by treatment with of SE against at RA in HEPM cells Fig. 3A: Apotracker staining (green) of HEPM cells after treatment with 100 μM at RA and 0.03% SE for 48 h. The nuclei were counterstained with Hoechst 33342 (blue). Scale bar, 50 μm. Fig. 3B: Immunoblotting of HEPM cells after treatment with 100 μM at RA and 0.03% SE for 48 h. β-actin was served as an internal control. *p < 0.05 and ***p < 0.001 versus at RA. at RA: all-trans -retinoic acid HEPM: human embryonic palatal mesenchymal SE: Sasa veitchii extract CCND1: cyclin D1

Article Snippet: Rabbit anti-cleaved caspase-3 polyclonal antibody (1:2,500 dilution; Cell Signaling Technology, Beverly, MA), mouse anti-cyclin D1 (CCND1) monoclonal antibody (1:1,000 dilution; Santa Cruz Biotechnology, Dallas, TX), mouse anti-CCNA (1:1,000 dilution; Santa Cruz Biotechnology), anti-mouse CCNB (1:1,000 dilution; Santa Cruz Biotechnology), anti-mouse CCNE (1:1,000 dilution; Santa Cruz Biotechnology), and anti-mouse β-actin monoclonal antibody (1:2,500 dilution; MBL, Aichi, Japan) were used as primary antibodies for immunoblotting.

Techniques: Staining, Western Blot, Control

Journal: Nagoya Journal of Medical Science

Article Title: Protective effect of Sasa veitchii extract against all-trans-retinoic acid-induced inhibition of proliferation of cultured human palate cells

doi: 10.18999/nagjms.86.2.223

Figure Lengend Snippet: Proposed mechanism of SE against at RA-induced cell proliferation inhibition ERBB2: Erb-B2 receptor tyrosine kinase 2 JADE1 : jade family PHD finger 1 CCND1: cyclin D1

Article Snippet: Rabbit anti-cleaved caspase-3 polyclonal antibody (1:2,500 dilution; Cell Signaling Technology, Beverly, MA), mouse anti-cyclin D1 (CCND1) monoclonal antibody (1:1,000 dilution; Santa Cruz Biotechnology, Dallas, TX), mouse anti-CCNA (1:1,000 dilution; Santa Cruz Biotechnology), anti-mouse CCNB (1:1,000 dilution; Santa Cruz Biotechnology), anti-mouse CCNE (1:1,000 dilution; Santa Cruz Biotechnology), and anti-mouse β-actin monoclonal antibody (1:2,500 dilution; MBL, Aichi, Japan) were used as primary antibodies for immunoblotting.

Techniques: Inhibition

Journal: Frontiers in Immunology

Article Title: The bone marrow endothelial progenitor cell response to septic infection

doi: 10.3389/fimmu.2024.1368099

Figure Lengend Snippet: (A) Fold changes in bone marrow BrdU + and BrdU − LKS VEGFR2 + cells in mice 24 h following i.v. challenge with E. coli (~1 × 10 8 CFUs/mouse). N = 5. Data are mean ± SEM. **p < 0.01 vs. the correspondent saline group. (B, C) Upregulation of cyclin D1 expression by marrow lin − c-kit + cells 24 h following i.v. challenge with E. coli (~5 × 10 7 CFUs/mouse). Data are mean ± SEM. MCF, mean channel fluorescence intensity. N = 5. **p < 0.01 vs. the saline group. (D) Increase in SP1 + cells in bone marrow LKS and lin − c-kit + cell subpopulations 18 h post i.v. challenge with E. coli (~5 × 10 7 CFUs/mouse). N = 5. Data are mean ± SEM. *p < 0.05 compared to the corresponding saline group; **p < 0.01 compared to the corresponding saline group. (E1, E2) Representative images of 5-week vasculogenic activity in implanted Matrigel plugs containing bone marrow lin − c-kit + cells from donor GFP + mice challenged with i.v. saline and heat-inactivated E. coli (~1 × 10 8 CFUs/mouse), respectively, for 24 h. (E2a) An amplified image in the area framed by the yellow square in (E2) .

Article Snippet: Permeabilized cells were incubated with 10 µg/ml of fluorochrome-conjugated anti-human/mouse cyclin D1 antibody (Clone DCS-6, Thermo Fisher Scientific, Inc.) or anti-human/mouse SP1 antibody (clone E-3, Santa Cruz Biotechnology, Inc.) in the dark for 20 min at room temperature.

Techniques: Saline, Expressing, Fluorescence, Activity Assay, Amplification