gpx4 activity  (Elabscience Biotechnology)

Journal: iScience

Article Title: NUP62 promotes breast cancer progression and inhibits ferroptosis by stabilizing NRF2 in a KEAP1-dependent way

doi: 10.1016/j.isci.2026.115484

Figure Lengend Snippet: NUP62 stabilizes the NRF2 protein by decreasing its degradation (A) The expression levels of antioxidant and ferroptosis-related genes were quantified by RT-qPCR in indicated BC cells. (B) SLC7A11 and GPX4 protein expressions were measured by IF staining in BC cells with NUP62 overexpression. Scale bars, 10 μm. (C) SLC7A11 and GPX4 protein expressions were measured by IF staining in BC cells with NUP62 knockdown. Scale bars, 10 μm. (D) GPX4 enzyme activity was measured in BC cells after NUP62 knockdown or overexpression. (E) mRNA levels of NRF2 in the BC cells with NUP62 knockdown or overexpression. (F) Protein levels of NRF2, SLC7A11, and GPX4 in the BC cells with NUP62 knockdown or overexpression. (G) BC cells with NUP62 knockdown or controls were treated with MG132 (10 μM) for 4 h, and NRF2 protein levels were analyzed by western blotting. (H) NRF2 protein half-life of BC cells with NUP62 overexpression or knockdown was evaluated using western blotting. (I) NRF2 ubiquitination levels of BC cells with NUP62 overexpression or knockdown. (J) NUP62 knockdown inhibited NRF2 nuclear translocation in BC cells, as evidenced by IF microscopy and western blotting. Scale bars, 10 μm. (K) NUP62 overexpression facilitated NRF2 nuclear translocation in BC cells, as evidenced by IF microscopy and western blotting. Scale bars, 10 μm. Data are presented as the mean ± SD. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001.

Article Snippet: GPX4 activity was quantified using a GPX4-specific Activity Assay Kit (Elabscience, E-BC-K883-M) following manufacturer's protocol.

Techniques: Expressing, Quantitative RT-PCR, Staining, Over Expression, Knockdown, Activity Assay, Western Blot, Ubiquitin Proteomics, Translocation Assay, Microscopy

Journal: iScience

Article Title: NUP62 promotes breast cancer progression and inhibits ferroptosis by stabilizing NRF2 in a KEAP1-dependent way

doi: 10.1016/j.isci.2026.115484

Figure Lengend Snippet: Eribulin destabilizes NRF2 protein and inhibits its nuclear localization (A) The protein levels of NRF2 in eribulin-treated BC cells with or without NUP62 knockdown. (B) NRF2 protein half-life of eribulin-treated BC cells with or without NUP62 knockdown, evaluated using western blotting. (C) NRF2 ubiquitination levels of eribulin-treated BC cells with or without NUP62 knockdown. (D) Nucleus-cytoplasm distribution of NUP62 in indicated BC cells, as evidenced by western blotting. (E) Nucleus-cytoplasm distribution of NUP62 in indicated BC cells, as evidenced by IF microscopy. (F) SLC7A11 protein expression , measured by IF staining in indicated BC cells. (G) GPX4 protein expression, measured by IF staining in indicated BC cells. Data are presented as the mean ± SD. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001.

Article Snippet: GPX4 activity was quantified using a GPX4-specific Activity Assay Kit (Elabscience, E-BC-K883-M) following manufacturer's protocol.

Techniques: Knockdown, Western Blot, Ubiquitin Proteomics, Microscopy, Expressing, Staining

Journal: iScience

Article Title: NUP62 promotes breast cancer progression and inhibits ferroptosis by stabilizing NRF2 in a KEAP1-dependent way

doi: 10.1016/j.isci.2026.115484

Figure Lengend Snippet: Eribulin or NUP62 knockdown inhibits tumor growth in vivo (A) The volume and size of subcutaneous tumors in mice with or without eribulin treatment. (B) The volume and size of subcutaneous tumors in mice with NUP62 knockdown. (C) The volume and size of subcutaneous tumors in mice after NRF2 knockdown in NUP62-overexpressing MDA-MB-231 cells. (D–F) Representative IHC staining images of Ki-67, NUP62, NRF2, and GPX4 in tumor tissue sections of each group. (G–I) Lipid peroxidation and GSH/GSSG ratio were measured in tumor tissue of each group. (J) A schematic model showing that NUP62 protects BC cells against ferroptosis by sustaining NRF2 signaling activation. Data are presented as the mean ± SD. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001.

Article Snippet: GPX4 activity was quantified using a GPX4-specific Activity Assay Kit (Elabscience, E-BC-K883-M) following manufacturer's protocol.

Techniques: Knockdown, In Vivo, Immunohistochemistry, Activation Assay

Journal: Redox Biology

Article Title: Copper deprivation reprograms antioxidant defense to suppress ferroptosis via SLC7A11

doi: 10.1016/j.redox.2026.104130

Figure Lengend Snippet: Copper deprivation induced by SLC31A11 knockdown triggers the upregulation of SLC7A11. ( A ) Confocal imaging of Cu-probe and Mito-tracker in the NC and SLC31A1 knockdown AsPC-1 cells. Hoechst (blue) is used as a nuclear counterstain. Mean ± SD, n = 3. Statistical significance was determined using a two-way ANOVA test. Scale bar: 2 μm. ( B ) Total Fe level in the NC and SLC31A1 knockdown AsPC-1, MiaPaCa-2, and CFPAC-1 cells. Mean ± SD, n = 3. Statistical significance was determined using a one-way ANOVA test. ( C ) Volcano plot of gene expression (the SLC31A1 knockdown [shRNA#2] versus the control; log2(fold change) ≥1; p < 0.05 between SLC31A1 knockdown and NC AsPC-1 cells. ( D ) GO analysis of differentially expressed genes between SLC31A1 knockdown (shRNA#2) and NC AsPC-1 cells. ( E-G ) Western blot analysis of the indicated protein levels in the NC and SLC31A1 knockdown AsPC-1 (E), MiaPaCa-2 (F), and CFPAC-1 (G) cells. ( H and I ) GPX4 (H) or FSP-1 activity (I) was tested in the NC and SLC31A1 knockdown AsPC-1 cells. Mean ± SD, n = 3. Statistical significance was determined using a one-way ANOVA test. ( J ) qPCR analysis of SLC31A1 and SLC7A11 gene expression in NC and SLC31A1 knockdown AsPC-1, MiaPaCa-2, and CFPAC-1 cells. Mean ± SD, n = 3. Statistical significance was determined using a one-way ANOVA test. ( K ) qPCR analysis of SLC31A1 and SLC7A11 gene expression in NC and SLC31A1 knockdown AsPC-1, MiaPaCa-2, and CFPAC-1 cells. Mean ± SD, n = 3. Statistical significance was determined using a one-way ANOVA test. ( L ) GSH level in the NC and SLC31A1 knockdown AsPC-1, MiaPaCa-2, and CFPAC-1 cells. Mean ± SD, n = 3. Statistical significance was determined using a one-way ANOVA test. ( M ) Cystine uptake level in the NC and SLC31A1 knockdown AsPC-1, MiaPaCa-2, and CFPAC-1 cells. Mean ± SD, n = 3. Statistical significance was determined using a one-way ANOVA test.

Article Snippet: Glutathione Peroxidase 4 (GPX4) Activity Assay Kit , Elabscience , E-BC-K883-M.

Techniques: Knockdown, Imaging, Gene Expression, shRNA, Control, Western Blot, Activity Assay