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NRF2 knockdown potentiates radiation-induced immunogenic cell death (ICD) in esophageal squamous cell carcinoma cells.(A, D) Western blot analysis of NRF2, Calreticulin (CRT), and GPX2 protein expression in KYSE150 (A) and KYSE30 (D) cells following irradiation (0, 6, 9 Gy). (B, E) Quantification <t>of</t> <t>extracellular</t> <t>ATP</t> secretion in the culture supernatants of KYSE150 (B) and KYSE30 (E) cells at 24 and 48 hours post-irradiation.(C, F) ELISA analysis of HMGB1 release in the supernatants of KYSE150 (C) and KYSE30 (F) cells 24 hours after irradiation with the indicated doses.(G, H) Flow cytometric analysis of cell surface CRT exposure in KYSE150 (G) and KYSE30 (H) cells. (I–L) Representative immunofluorescence images and quantitative analysis of NRF2 (red) and CRT (green) expression in KYSE30 (I, J) and KYSE150 (K, L) cells. Nuclei were counterstained with DAPI (blue). Scale bars = 20 μm. For each group, n = 3 independent biological replicates (slides) were used. Three non-overlapping fields of view were randomly selected from each slide for image acquisition and analysis. Quantitative data are presented as mean ± standard deviation (SD) or standard error of the mean (SEM).
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NRF2 knockdown potentiates radiation-induced immunogenic cell death (ICD) in esophageal squamous cell carcinoma cells.(A, D) Western blot analysis of NRF2, Calreticulin (CRT), and GPX2 protein expression in KYSE150 (A) and KYSE30 (D) cells following irradiation (0, 6, 9 Gy). (B, E) Quantification of extracellular ATP secretion in the culture supernatants of KYSE150 (B) and KYSE30 (E) cells at 24 and 48 hours post-irradiation.(C, F) ELISA analysis of HMGB1 release in the supernatants of KYSE150 (C) and KYSE30 (F) cells 24 hours after irradiation with the indicated doses.(G, H) Flow cytometric analysis of cell surface CRT exposure in KYSE150 (G) and KYSE30 (H) cells. (I–L) Representative immunofluorescence images and quantitative analysis of NRF2 (red) and CRT (green) expression in KYSE30 (I, J) and KYSE150 (K, L) cells. Nuclei were counterstained with DAPI (blue). Scale bars = 20 μm. For each group, n = 3 independent biological replicates (slides) were used. Three non-overlapping fields of view were randomly selected from each slide for image acquisition and analysis. Quantitative data are presented as mean ± standard deviation (SD) or standard error of the mean (SEM).

Journal: Journal of Translational Medicine

Article Title: Nrf2 deficiency converts the ESCC microenvironment into an immunologically active state via the GPX2-ICD-DC signaling path

doi: 10.1186/s12967-026-08323-7

Figure Lengend Snippet: NRF2 knockdown potentiates radiation-induced immunogenic cell death (ICD) in esophageal squamous cell carcinoma cells.(A, D) Western blot analysis of NRF2, Calreticulin (CRT), and GPX2 protein expression in KYSE150 (A) and KYSE30 (D) cells following irradiation (0, 6, 9 Gy). (B, E) Quantification of extracellular ATP secretion in the culture supernatants of KYSE150 (B) and KYSE30 (E) cells at 24 and 48 hours post-irradiation.(C, F) ELISA analysis of HMGB1 release in the supernatants of KYSE150 (C) and KYSE30 (F) cells 24 hours after irradiation with the indicated doses.(G, H) Flow cytometric analysis of cell surface CRT exposure in KYSE150 (G) and KYSE30 (H) cells. (I–L) Representative immunofluorescence images and quantitative analysis of NRF2 (red) and CRT (green) expression in KYSE30 (I, J) and KYSE150 (K, L) cells. Nuclei were counterstained with DAPI (blue). Scale bars = 20 μm. For each group, n = 3 independent biological replicates (slides) were used. Three non-overlapping fields of view were randomly selected from each slide for image acquisition and analysis. Quantitative data are presented as mean ± standard deviation (SD) or standard error of the mean (SEM).

Article Snippet: Extracellular ATP release was measured using an ATP Detection Kit (MCE HY-K0314) according to the manufacturer’s instructions.

Techniques: Knockdown, Western Blot, Expressing, Irradiation, Enzyme-linked Immunosorbent Assay, Immunofluorescence, Standard Deviation