Journal: Translational Oncology

Article Title: NF-κB activation as a pro-survival signal from pharmacological inhibition of pyruvate dehydrogenase kinase 1 in non-small-cell lung carcinoma cell models

doi: 10.1016/j.tranon.2026.102681

Figure Lengend Snippet: The activation of NF-κB serves as a survival mechanism to rescue 64 -induced inhibition of NSCLC cell growth and apoptosis. NCI-H1975 and NCI-H1650 cells were treated with 64 alone or in combination with JSH-23. a . Western blotting of P65 in the cytoplasm and nuclear was conducted under treatment with 64 and JSH-23 for 24 h. b . MTT assay was conducted to determine cell viability under treatment with 64 and JSH-23 for 24 h. c . The cell viability inhibition by 64 (0.5µM, 1µM, 2µM, 3µM, 4µM, 8µM) and JSH-23(2.5µM, 5µM, 10µM, 15µM, 20μΜ, 40μΜ) was also evaluated, as was the combined effect of the two compounds. d . The CI (Combination Index) values were calculated using the software Biosoft CalcuSyn to determine the impact of individual combined concentrations of 64 and JSH-23 (fraction affected (Fa)) accordance with cell viability. e . JC-1 assay to assess MMP under treatment with 64 and JSH-23 for 4 h. f . Annexin V-PI double staining assay to evaluate cell apoptosis under treatment with 64 and JSH-23 for 4 h. The data were obtained from three independent experiments with n = 3 and were presented as mean ± SD. Statistical significance between compared groups was indicated by * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: P38 MAPK inhibitor SB203580 (S1076) and NF-κB inhibitor JSH-23 (S7351) were purchased from SelleckChem (Houston, TX, USA).

Techniques: Activation Assay, Inhibition, Western Blot, MTT Assay, Software, Double Staining

Journal: Translational Oncology

Article Title: NF-κB activation as a pro-survival signal from pharmacological inhibition of pyruvate dehydrogenase kinase 1 in non-small-cell lung carcinoma cell models

doi: 10.1016/j.tranon.2026.102681

Figure Lengend Snippet: The activation of NF-κB pathway caused by 64 affecting the expressions levels key apoptotic proteins in NSCLC cells. The NSCLC cell lines NCI-H1975 and NCI-H1650 were treated with 64 alone or in combination with JSH-23 for 24 h. a . The expression levels of CytC in the mitochondria and cytoplasm were detected by western blotting. b . The histograms showing the ratios between cytoplasmic CytC to mitochondrial CytC. c . The expressions of apoptotic-related proteins were detected by western blotting. d . The histogram analysis for the expression of Bax and Bcl-2, as well as the ratios between the cleaved form (c-cas3 and c-PARP) and the total form (cas3 and PARP). e . The expression of proteins in the JNK pathway were detected by western blotting. f . The histogram analysis for the ratios of phosphorylated form (p-JNK and p-c-Jun) to total form (JNK and c-Jun). The data were obtained from three independent experiments with n = 3 and were presented as mean ± SD. The statistical significance between the compared groups was indicated by * P < 0.05, ** P < 0.01, *** P <0.001.

Article Snippet: P38 MAPK inhibitor SB203580 (S1076) and NF-κB inhibitor JSH-23 (S7351) were purchased from SelleckChem (Houston, TX, USA).

Techniques: Activation Assay, Expressing, Western Blot

Journal: Frontiers in Immunology

Article Title: Dysregulated hyaluronan metabolism drives inflammation and angiogenesis in proliferative diabetic retinopathy

doi: 10.3389/fimmu.2026.1724199

Figure Lengend Snippet: Human retinal Müller glial cells were left untreated or treated with ultra-low molecular weight hyaluronan (ULMW-HA) (50 µg/mL) for 24 (h) (A) Protein expression of phospho-ERK1/2 and phospho-NFκB in cell lysates was determined by Western blot analysis. Levels of high mobility group box-1 (HMGB1) were quantified in the culture media by ELISA. Results are expressed as mean ± standard deviation from three different experiments each performed in triplicate (*p < 0.05; independent t-test). (B) Human retinal Müller glial cells were left untreated or treated with ULMW-HA, ULMW-HA plus BAY11-7085 (5 µM) or (C) ULMW-HA plus U-0126 (5 µM). Levels of vascular endothelial growth factor (VEGF), angiopoietin and monocyte chemotactic protein-1 (MCP-1/CCL2) were quantified in the culture media by ELISA. Results are expressed as mean ± standard deviation or standard error of mean from three different experiments each performed in triplicate. One-way ANOVA and independent t-test were used for comparisons between three groups and two groups, respectively. *p < 0.05 compared with values obtained from untreated cells; #p < 0.05 compared with ULMW-HA plus BAY11–7085 or U-0126 treated cells. (D, E) Human retinal Müller glial cells were left untreated or treated with high glucose (HG) (25 mM), cobalt chloride (CoCl 2 ) (300 µM) or tumor necrosis factor-α (TNF-α) (5 ng/mL) with or without apigenin (10 µg/mL) for 24 (h) For HG treatment, cultures containing 25 mM mannitol were used as a control. Levels of monocyte chemotactic protein-1 (MCP-1/CCL2) (D) and vascular endothelial growth factor (VEGF) (E) were quantified in the culture media by ELISA. The results are expressed as mean ± standard deviation from three different experiments each performed in triplicate. One-way ANOVA and independent t-test were used for comparisons between three and two groups, respectively. *p < 0.05 compared with values obtained from control cells. #p < 0.05 compared with values obtained from stimulated cells.

Article Snippet: Additionally, overnight starved Müller cells were treated with hyaluronan (ULMW-HA at 50 μg/ml, Cat No GLR003, R&D Systems) in the absence or presence of 1h pretreatment with the nuclear factor-kappa B (NF-κB) inhibitor BAY11-7085 (5 μM, Cat No sc-202490, Santa Cruz Biotechnology Inc., Santa Cruz, CA USA), or the ERK1/2 inhibitor U0126 (5 μM, Cat No sc-222395A, Santa Cruz Biotechnology Inc.) or the combination of the NF-κB inhibitor BAY11–7085 and the ERK1/2 inhibitor U0126 for 16 hours.

Techniques: Molecular Weight, Expressing, Western Blot, Enzyme-linked Immunosorbent Assay, Standard Deviation, Control