Journal: Redox Biology

Article Title: ALDH1L2 induces resistance to chemotherapy in small cell lung cancer by inhibiting ferroptosis

doi: 10.1016/j.redox.2026.104098

Figure Lengend Snippet: ALDH1L2 promotes SCLC chemoresistance by negatively regulating the hyperoxidized PRDX3 and PRDX3 dimer content in the plasma membrane. (A) Immunofluorescence detection of the colocalization of ALDH1L2 and PRDX3 in SCLC cells. The cell nuclei are labeled with blue fluorescence, the ALDH1L2 protein is labeled with green fluorescence, and the PRDX3 protein is labeled with red fluorescence. Scale bar: 20 μm. (B) Coimmunoprecipitation assays revealed the interaction of the ALDH1L2 protein with the PRDX3 protein in chemoresistant SCLC cells. (C) Coimmunoprecipitation assays revealed the interaction of the TRX2 protein with the ALDH1L2 protein and the PRDX3 protein in chemoresistant SCLC cells. (D-E) The reaction of PRDX3 with hydroperoxides was analyzed by immunoblotting. Bar plots show the results of the semiquantitative analysis of the gray values of the protein bands by ImageJ software. (F–H) Analysis of the effect of ALDH1L2 knockdown on the content of hyperoxidized PRDXs protein in each fraction of H69AR and H446DDP cells. The results under no treatment, cisplatin treatment (10 μg/mL for 24 h) and erastin treatment (20 μM for 72 h) are shown in the graph. Bar plots show the results of the semiquantitative analysis of the gray values of the protein bands by ImageJ software. (I) Coimmunoprecipitation assays revealed that hyperoxidized PRDXs protein could be immunoprecipitated by anti-PRDX3 antibody. (J-N) Analysis of the effect of ALDH1L2 knockdown on the content of oxidized PRDX3 dimers in each fraction of H69AR and H446DDP cells. The results under no treatment, cisplatin treatment (10 μg/mL for 24 h) and erastin treatment (20 μM for 72 h) are shown in the graph. LE, long exposure; SE, short exposure. Bar plots show the results of the semiquantitative analysis of the gray values of the protein bands by ImageJ software. The data are expressed as mean ± standard deviation (n = 3). ns, not significant; ∗, p < 0.05; ∗∗, p < 0.01; ∗∗∗, p < 0.001; ∗∗∗∗, p < 0.0001.

Article Snippet: The purified PRDX3 protein was obtained from MedChemExpress (HY– P71147 , MCE, USA).

Techniques: Clinical Proteomics, Membrane, Immunofluorescence, Labeling, Fluorescence, Western Blot, Software, Knockdown, Immunoprecipitation, Standard Deviation

Journal: Redox Biology

Article Title: ALDH1L2 induces resistance to chemotherapy in small cell lung cancer by inhibiting ferroptosis

doi: 10.1016/j.redox.2026.104098

Figure Lengend Snippet: PRDX3 contributes to the ALDH1L2-mediated chemoresistance program in SCLC. (A-D) RT-qPCR was used to verify the knockdown and overexpression efficiency of PRDX3 in SCLC cells at the mRNA level. (E-L) Immunoblotting was performed to validate the knockdown and overexpression efficiency of PRDX3 in SCLC cells at the protein level. Bar plots show the results of the semiquantitative analysis of the gray values of the protein bands by ImageJ software. (M − P) CCK-8 assay to determine the role of PRDX3 in the regulation of chemoresistance elicited by ALDH1L2 in SCLC cells. IC50, half maximal inhibitory concentration. The data are expressed as mean ± standard deviation (n = 3). ns, not significant; ∗, p < 0.05; ∗∗, p < 0.01; ∗∗∗, p < 0.001; ∗∗∗∗, p < 0.0001.

Article Snippet: The purified PRDX3 protein was obtained from MedChemExpress (HY– P71147 , MCE, USA).

Techniques: Quantitative RT-PCR, Knockdown, Over Expression, Western Blot, Software, CCK-8 Assay, Concentration Assay, Standard Deviation

Journal: Redox Biology

Article Title: ALDH1L2 induces resistance to chemotherapy in small cell lung cancer by inhibiting ferroptosis

doi: 10.1016/j.redox.2026.104098

Figure Lengend Snippet: The PRDX3 inhibitor thiostrepton synergizes with chemotherapy to suppress tumor growth in SCLC. (A) CCK-8 assay to determine the IC50 values of thiostrepton in human normal bronchial epithelial BEAS-2B cells and SCLC cells (n = 3). (B–C) Verification of the inhibitory effect of thiostrepton on PRDX3 in SCLC cells by immunoblotting. Bar plots show the results of the semiquantitative analysis of the gray values of the protein bands by ImageJ software (n = 3). (D) The growth of tumors in the orthotopic SCLC model was recorded with the live animal imaging system (n = 5). (E) Line chart demonstrating the quantitative bioluminescence intensities for each group. (F) Growth curves of xenografted tumors derived from H69AR cells under different treatments are shown (n = 5). The tumor volume was calculated as follows: (longest diameter) × (shortest diameter) 2 /2. (G) Xenografted tumors derived from H69AR cells under different treatments were removed and photographed after the mice were euthanized. (H) Weights of xenografted tumors derived from H69AR cells under different treatments are shown. IC50, half maximal inhibitory concentration. The data are expressed as mean ± standard deviation. ns, not significant; ∗, p < 0.05; ∗∗, p < 0.01; ∗∗∗, p < 0.001; ∗∗∗∗, p < 0.0001.

Article Snippet: The purified PRDX3 protein was obtained from MedChemExpress (HY– P71147 , MCE, USA).

Techniques: CCK-8 Assay, Western Blot, Software, Imaging, Derivative Assay, Concentration Assay, Standard Deviation