Fig 1: Analysis of candidate biomarkers mRNA expression.DDX39A (A), HMGA1 (B), HMGA2 (C), HOXC9 (D), and PBX1 (E). Relative gene expression was determined in 6 patient-derived and 2 reference cell lines after 7 days of treatment with natural (ATRA, 9-cis-RA, 13-cis-RA) and synthetic (BEX, 4 HPR) retinoids administered at a 1 μM concentration. Experiments were repeated in triplicate. Representative gel electrophoresis images captured using a UV transilluminator are shown for each sample, including HSP90AB1 as a reference gene.
Fig 2: Representative IHC analysis of putative biomarkers in tumor samples.Representative expressions of DDX39A, HMGA1, HMGA2, HOXC9, and PBX1 in the tumor samples No. 2 (corresponding to the NBL-13 retinoid-sensitive cell line) and No. 18 (corresponding to the NBL-36 retinoid-resistant cell line). Representative expressions of these markers in positive control samples are also given for comparison. Original magnification, 200×.
Fig 3: Analysis of candidate biomarkers protein expression.(A) DDX39A–49 kDa; (B) HMGA1–18 kDa; (C) HMGA2–18 kDa; (D) HOXC9–29 kDa; and (E) PBX1–46 kDa. Relative protein expression was determined in 6 patient-derived and 2 established NBL cell lines after treatment with natural (ATRA, 9-cis-RA, 13-cis-RA) and synthetic (BEX, 4-HPR) retinoids administered at a 1 μM concentration for 7 days. Images of the representative immunoblot captured using X-ray film are shown for each sample, including GAPDH as a reference protein. Experiments were repeated in triplicate.
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