Fig 2: ONC201/ONC206 treatment induces differential expression of tumorigenic proteins in IMR-32 NB cells. IMR-32 cells treated with either ONC201 (5 μM) or ONC206 (0.5 μM) exhibited significant reduction of tumorigenic protein expression compared with vehicle-treated cells as assessed using Western blot analyses. (A) Western blot quantitative analysis revealed significant downregulation in the protein expression of tumorigenic NMYC, FABP5, and HMGA1 as well as stem-markers Oct-4 and Sox-2, 48 h after drug treatment. (B) Cleaved caspase-3, cleaved-PARP1, and γH2AX were upregulated at 48 and 72 h post-drug treatment. Blots without GAPDH loading control were quantitated using Bio-Rad's stain-free gel technology and normalized to the total protein loaded per lane. Experiments were conducted in triplicates and repeated three times. Data represent the mean ± the standard error of the mean (SEM) of multiple experiments. *p < 0.05; **p < 0.01; ***p < 0.001.

Fig 3: Alterations of protein S-glutathionylation modification in macrophages under oxidative conditions.a Schematic of the enrichment and quantitative analysis for SSG-modified Cys-peptides. b Enrichment of protein SSG and LC–MS/MS in BMDMs exposed to H2O2 (200 µM) for 0, 15 min. Gene ontology annotations for biological process of proteins with significant S-glutathionylation alterations found to be increased or decreased in any stage by >1.2-fold compared to control. The slice for metabolic process is slightly displaced from the rest of the pie chart. c Alterations in S-glutathionylation sites under exposure to H2O2 [shown as fold changes of protein S-glutathionylation (H2O2 treated/control). Gray, identified protein sites; blue, metabolism-related differential proteins; orange, FABP5]. d mRNA levels of Ptbp1, Otub1, Ctsz, Gapdh, and Fabp5 in BMDMs as evaluated by qPCR, n = 3 biologically independent samples over 3 independent experiments, data are shown as mean ± SEM and analyzed with a 95% confidence interval, P < 0.0001, one-way ANOVA is followed by Tukey’s post-hoc test. Source data are provided as a Source Data file.

Fig 4: PPARβ/δ signaling might not be involved in FABP5-mediated growth promotion of PCa cells(A) PPARβ/δ mRNA expression levels in PNT2, DU-145, PC-3 and PC-3M. Relative mRNA levels were measured by qPCR. Results are means ± S.D. for three independent experiments. (B) PNT2 cells were transfected with pCI-neo or pCI-neo/FABP5. At 48 h after transfection, transfected cells were treated with DMSO or GW0742 (2 μM) for 24 h. mRNA levels of FABP5, PDPK1, ADRP and ILK were measured by qPCR and normalized to those of 18S rRNA. Results are means ± S.D. for three independent experiments. (C) PC-3 cells were transfected with control siRNA (20 nM) or siFABP5 (20 nM). At 48 h after transfection, cells were treated with DMSO or GW0742 (2 μM) for 24 h. FABP5, PDPK1, ADRP and ILK mRNA levels were measured by qPCR and normalized to those of 18S rRNA. Results are means ± S.D. for three independent experiments. (D) Phosphorylation of Akt (S473) in siFABP5 transfected DU-145 and PC-3 was detected by western blot analysis. We used insulin (as positive control of AKT phosphorylation, 1 μg/ml, 15 min) and wortmannin (inhibitor of phosphoinositide 3-kinases, as negative control of AKT phosphorylation, 0.5 μM, 30 min). Results shown are representative of three independent experiments. **P < 0.01, one-way ANOVA followed by Tukey's test (B and C), N.S. = Not significant.

Fig 5: Nuclear localization of FABP5 promotes cell proliferation(A) Western blot analysis of FABP5 and NES-FABP5 in PNT2 cells transfected with pCI-neo/FABP5, NES-FABP5 or pCI-neo (e.v., empty vector). Results shown are representative of three independent experiments. (B) Localization of FABP5 and NES-FABP5 in FABP5 or NES-FABP5 overexpressing PNT2 cells. Transfected cells were treated with ethanol or oleic acid (10 μM) for 30 min. (C) Cell growth of FABP5- and NES-FABP5-transfected PNT2 cells. Cells were counted at the indicated times. Results are means ± S.D. for three independent experiments. **P < 0.01, two-way ANOVA followed by Tukey's test.