Journal: Journal of Translational Medicine

Article Title: Effect and underlying mechanism of a photochemotherapy dual-function nanodrug delivery system for head and neck squamous cell carcinoma

doi: 10.1186/s12967-024-05855-8

Figure Lengend Snippet: PN and PND exerted antitumour effect on HNSCC in vivo. A The procedure for the xenograft tumour model in nude mice. B The body weight of nude mice was measured every 2 days. C Tumour volume was measured every 2 days. D The tumour inhibition rate was calculated after death of nude mice. E Morphological images of xenograft tumours. F Tumour weight at the end of the experiment. G HE staining of transplanted tumours, heart, liver, lung, and kidney in nude mice. Scale bar: 50 μm. H-I IF staining of Bax, Bcl-2, E-cadherin, N-cadherin and Vimentin, p-ASK1, p-JNK, c-Fos and c-Jun, along with quantitative analysis of staining intensity. Scale bar: 50 μm. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001. All data are shown as the mean ± SD from three independent experiments

Article Snippet: The primary antibodies used in this study against B-cell CLL/lymphoma 2 (Bcl-2), BCL2 associated X (Bax), E-cadherin (E-ca), N-cadherin (N-ca), Vimentin, and phosphorylated ASK1 (p-ASK1) and the secondary antibody horseradish peroxidase-conjugated affinipure goat anti-rabbit IgG were all purchased from Proteintech (Wuhan, China).

Techniques: In Vivo, Inhibition, Staining

Journal: International Journal of Molecular Sciences

Article Title: Copper Oxide Nanoparticles Induce Pulmonary Inflammation and Exacerbate Asthma via the TXNIP Signaling Pathway

doi: 10.3390/ijms252111436

Figure Lengend Snippet: Effects of CuO NP treatment on the protein expression of TXNIP, p-ASK1, total ASK1 (t-ASK1), Bax, Bcl-2, and cleaved caspase-3. ( A ) Representative figure of TXNIP expression on lung tissue via ICH (×200, Bar = 60 nm). ( B ) Expression value of TXNIP. ( C ) Protein expression detected on the Western blot gels. ( D – I ) Relative densitometric values of each protein. Mice were necropsied 48 h after the last dose of CuO NPs. Data are indicated as mean ± SD ( n = 3) Significant differences from the control group are shown by the following symbols: * p < 0.05, ** p < 0.01, and *** p < 0.001; “ns” indicates not significant ( p > 0.05).

Article Snippet: The following primary antibodies and dilutions were used: TXNIP (1:1000; Novus Biologicals); phosphorylated ASK1 (p-ASK1, 1:1000; Cell Signaling Technology); total ASK1 (t-ASK1, 1:1000; Abcam, Cambridge, UK); Bax (1:1000; Cell Signaling Technology); Bcl-2 (1:1000; Cell Signaling Technology); cleaved caspase-3 (1:1000; Cell Signaling Technology); and β-actin (1:1000; Cell Signaling Technology).

Techniques: Expressing, Western Blot, Control

Journal: International Journal of Molecular Sciences

Article Title: Copper Oxide Nanoparticles Induce Pulmonary Inflammation and Exacerbate Asthma via the TXNIP Signaling Pathway

doi: 10.3390/ijms252111436

Figure Lengend Snippet: Effects of CuO NP treatment on the TXNIP signaling pathway in NCI-H292 cells. ( A ) Protein expression detected on the Western blot gels. ( B – G ) Relative densitometric values of each protein. ( H ) Representative figure for TXNIP and p-ASK1 expression by double-immunofluorescence staining in cells treated with hydrogen peroxide (100 μM), CuO NPs (2.0 μg/mL), and an untreated control (Bar = 10 μm). The cells were treated with a free medium, hydrogen peroxide, or CuO NPs (0.25, 0.5, 1.0, and 2.0 μg/mL) for 12 h. Data are indicated as mean ± SD ( n = 3). Significant differences from the control group are shown by the following symbols: * p < 0.05, ** p < 0.01, and *** p < 0.001; “ns” indicates not significant ( p > 0.05).

Article Snippet: The following primary antibodies and dilutions were used: TXNIP (1:1000; Novus Biologicals); phosphorylated ASK1 (p-ASK1, 1:1000; Cell Signaling Technology); total ASK1 (t-ASK1, 1:1000; Abcam, Cambridge, UK); Bax (1:1000; Cell Signaling Technology); Bcl-2 (1:1000; Cell Signaling Technology); cleaved caspase-3 (1:1000; Cell Signaling Technology); and β-actin (1:1000; Cell Signaling Technology).

Techniques: Expressing, Western Blot, Double Immunofluorescence Staining, Control

Journal: Oncology Reports

Article Title: ABT‑737 increases cisplatin sensitivity through the ROS‑ASK1‑JNK MAPK signaling axis in human ovarian cancer cisplatin‑resistant A2780/DDP cells

doi: 10.3892/or.2024.8781

Figure Lengend Snippet: ASK1 is required for ABT-737-induced JNK activation and apoptosis in A2780/DDP cells. (A) A2780/DDP cells were treated with cisplatin (5 µg/ml) alone or combined with ABT-737 (10 µM) for 24 h. Western blot analysis of p-ASK1 and ASK1 protein expression. (B) Quantitative analysis of p-ASK1/ASK1 protein levels from panel A. Data are presented as the mean ± SD of three independent experiments. *P<0.05 vs. the control group; ## P<0.01 vs. the cisplatin group. (C) A2780/DDP cells were treated with cisplatin (5 µg/ml) combined with ABT-737 (10 µM) or combined with ABT-737 (10 µM) and GS-4997 (5 µM) for 24 h. The cell viability was determined by MTT assays. Data are presented as the mean ± SD of three independent experiments. **P<0.01 vs. the control group; # P<0.05 vs. the cisplatin + ABT-737 group. (D) Hoechst staining analysis of nuclear morphology (scale bar, 10 µm). (E) Quantitative analysis of the Hoechst staining-positive cells. Data are presented as the mean ± SD of three independent experiments. **P<0.01 vs. the control group; ## P<0.01 vs. the cisplatin + ABT-737 group. (F) Cell apoptosis was detected by flow cytometry. (G) Quantitative analysis of the flow cytometry results. Data are presented as the mean ± SD of three independent experiments. **P<0.01 vs. the control group; ## P<0.01 vs. the cisplatin + ABT-737 group. (H) Western blot analysis of p-ASK1, ASK1, p-JNK and JNK protein expression. (I and J) Quantitative analysis of p-ASK1/ASK1 and p-JNK/JNK protein levels from panel H. Data are presented as the mean ± SD of three independent experiments. **P<0.01 vs. the control group; ## P<0.01 vs. the cisplatin + ABT-737 group. p-, phosphorylated.

Article Snippet: Antibodies against phosphorylated (p-) Ask1 (cat. no. 28846-1-AP; 1:1,000; rabbit), Ask1 (cat. no. 67072-1-Ig; 1:1,000; rabbit), p-P38 (cat. no. 28796-1-AP; 1:500; rabbit), P38 (cat. no. 14064-1-AP; 1:500; rabbit), p-Akt (cat. no. 66444-1-Ig; 1:2,000; mouse) and Akt (cat. no. 60203-2-Ig; 1:2,000; mouse) were purchased from Proteintech Group, Inc. Antibodies against caspase 3 (cat. no. 9662S; 1:2,000; rabbit), cleaved-caspase 3 (cat. no. 9661S; 1:1,000; rabbit), PARP (cat. no. 9542S; 1:1,000; rabbit), cleaved-PARP (cat. no. 9541S; 1:1,000; rabbit), p-JNK (cat. no. 4668S; 1:2,000; rabbit), JNK (cat. no. 9252S; 1:2,000; rabbit), p-ERK (cat. no. 4370S; 1:1,000; rabbit) and ERK (cat. no. 4695S; 1:1,000; rabbit) were purchased from Cell Signaling Technology, Inc.

Techniques: Activation Assay, Western Blot, Expressing, Control, Staining, Flow Cytometry

Journal: Oncology Reports

Article Title: ABT‑737 increases cisplatin sensitivity through the ROS‑ASK1‑JNK MAPK signaling axis in human ovarian cancer cisplatin‑resistant A2780/DDP cells

doi: 10.3892/or.2024.8781

Figure Lengend Snippet: ABT-737-induced production of reactive oxygen species mediates the activation of the ASK1-JNK signaling pathway and apoptosis of A2780/DDP cells. A2780/DDP cells were treated with cisplatin (5 µg/ml) combined with ABT-737 (10 µM) or combined with ABT-737 (10 µM) and NAC (100 µM) for 24 h. (A) Western blot analysis of p-ASK1, ASK1, p-JNK and JNK protein expression. (B and C) Quantitative analysis of (B) p-ASK1/ASK1 and (C) p-JNK/JNK protein levels from panel A. Data are presented as the mean ± SD of three independent experiments. (D) The cell viability was determined by MTT assays. Data are presented as the mean ± SD of three independent experiments. (E) Hoechst staining analysis of nuclear morphology (scale bar, 10 µm). (F) Quantitative analysis of the Hoechst staining-positive cells. Data are presented as the mean ± SD of three independent experiments. (G) Cell apoptosis was detected by flow cytometry. (H) Quantitative analysis of the flow cytometric results. Data are presented as the mean ± SD of three independent experiments. **P<0.01 vs. the control group; # P<0.05 and ## P<0.01 vs. the cisplatin + ABT-737 group. p-, phosphorylated.

Article Snippet: Antibodies against phosphorylated (p-) Ask1 (cat. no. 28846-1-AP; 1:1,000; rabbit), Ask1 (cat. no. 67072-1-Ig; 1:1,000; rabbit), p-P38 (cat. no. 28796-1-AP; 1:500; rabbit), P38 (cat. no. 14064-1-AP; 1:500; rabbit), p-Akt (cat. no. 66444-1-Ig; 1:2,000; mouse) and Akt (cat. no. 60203-2-Ig; 1:2,000; mouse) were purchased from Proteintech Group, Inc. Antibodies against caspase 3 (cat. no. 9662S; 1:2,000; rabbit), cleaved-caspase 3 (cat. no. 9661S; 1:1,000; rabbit), PARP (cat. no. 9542S; 1:1,000; rabbit), cleaved-PARP (cat. no. 9541S; 1:1,000; rabbit), p-JNK (cat. no. 4668S; 1:2,000; rabbit), JNK (cat. no. 9252S; 1:2,000; rabbit), p-ERK (cat. no. 4370S; 1:1,000; rabbit) and ERK (cat. no. 4695S; 1:1,000; rabbit) were purchased from Cell Signaling Technology, Inc.

Techniques: Activation Assay, Western Blot, Expressing, Staining, Flow Cytometry, Control