Journal: Cell Reports

Article Title: ALDH1A3-acetaldehyde metabolism potentiates transcriptional heterogeneity in melanoma

doi: 10.1016/j.celrep.2024.114406

Figure Lengend Snippet: ALDH1A3 High subpopulations promote melanoma drug resistance in vivo (A) Zebrafish BRAF V600E melanoma model (arrow indicates melanoma). BRAF V600E and GFP are expressed in the melanocyte lineage, by the mitfa promoter, and the zebrafish is mutant for p53. (B–D) ALDH High cells with low levels of mitfa:GFP in zebrafish melanoma (red circle). DEAB: negative control. (C) FACS of ALDH High and ALDH Low zebrafish melanoma cells. (D) RT-qPCR of sox2 , sox10 , and tfap2b expression in sorted ALDH High versus ALDH Low zebrafish melanoma cells ( n = 3 bio-replicates each with 3 technical replicates, multiple paired t test corrected with Holm-Sidak’s method). (E and F) Aldh1a3 in zebrafish model of BRAF inhibitor regression and recurrent disease. (E) BRAF V600E p53 mutant zebrafish with melanomas were fed with 200 mg/kg/day vemurafenib-containing food pellets leading to melanoma regression, drug resistance. and disease recurrence. (F) Aldh1a3 IHC in DMSO, BRAFi, and/or NAZ treatment-responding disease and in drug-resistant recurrent disease shows Aldh1a3 on-target efficacy of nifuroxazide. One-way ANOVA with Tukey’s test correction for multiple comparisons. Scale bar = 25 μm. ∗ p < 0.05; ∗∗∗ p < 0.001; ∗∗∗∗ p < 0.0001. (G and H) Zebrafish combination drug trial to target ALDH High cells. (G) Long-term drug-pellet treatment design. (H) Kaplan-Meier survival curves of zebrafish melanomas under different drug treatment shown in (G). Log rank tests, p < 0.001. See also Figure S6 .

Article Snippet: To perform drug treatment on adult zebrafish bearing melanoma, we produced fish bite-size drug pellets as described in our previous publication and fed single-housed individual fish with fish food agar pellets containing DMSO (Sigma Aldrich), vemurafenib (SelleckChem), and/or Nifuroxazide (Merck Millipore) once per day.

Techniques: In Vivo, Mutagenesis, Negative Control, Quantitative RT-PCR, Expressing

Journal: Cell Reports

Article Title: ALDH1A3-acetaldehyde metabolism potentiates transcriptional heterogeneity in melanoma

doi: 10.1016/j.celrep.2024.114406

Figure Lengend Snippet:

Article Snippet: To perform drug treatment on adult zebrafish bearing melanoma, we produced fish bite-size drug pellets as described in our previous publication and fed single-housed individual fish with fish food agar pellets containing DMSO (Sigma Aldrich), vemurafenib (SelleckChem), and/or Nifuroxazide (Merck Millipore) once per day.

Techniques: Recombinant, Control, Virus, Staining, Lysis, Protease Inhibitor, SYBR Green Assay, Red Blood Cell Lysis, Transfection, Infection, ALDH Detection Assay, Reverse Transcription, Sample Prep, Multiplex Assay, Purification, Bicinchoninic Acid Protein Assay, Software