Fig 2: Effects of RJW-58 in IL-10 expression via OLF-1. A, The cluster heat map reveals the results of microarray analysis. CTSS knockdown increases the mRNA levels of the olfactory receptor family. B, Western blot analysis of OR5B3, OR5I1, and OR5M3 proteins when SIM-A9 cells were treated RJW-58 (5 µM) for 24 h. C, OLF-1 disassociated with CBP by RJW-58 for 24 h treatment in SIM-A9 cells. D, The effect of RJW-58 for 24 h on OLF-1 binding to the IL-10 promoter region in SIM-A9 cells was analyzed using ChIP. E, RJW-58 treatment facilitated IL-10 promoter activity in SIM-A9. IL-10 promoters were identified using the promoter assay. ****P < 0.0001, ***P < 0.001, **P < 0.01, *P < 0.05. The statistical significance was determined using a two-tailed Student's t-test. Data are presented as the mean ± SEM.

Fig 3: The neuropathic effects of oxaliplatin in the Ctss+/+ mouse model. A, Flow analysis of ATF3 expression in ND 7/23 cell treated with control, oxaliplatin (1 µM), RJW-58 (5 µM), or a combination of oxaliplatin (1 µM) and RJW-58 (5 µM), respectively. B, The protocol of drug administration and behavioral tests in the mouse model. The basal levels of each behavioral assay were obtained before treatment. In the first week, PBS control (100 µL), oxaliplatin (3 mg/kg, 100 µL), and RJW-58 (25 mg/kg, 100 µL) were injected intraperitoneally five times per week for 2 weeks. Behavioral tests were performed weekly. C, Von Frey filament test to detect the paw withdrawal threshold to mechanical stimulus. The Y-axis is the normalized pressure from touch to paw withdrawal. D-E, Tail immersion test to assess thermal sensation. The Y-axis is the normalized latency from tail immersion to tail withdrawal. F, Locomotor activity test to detect allodynia. The Y-axis is the normalized moved distance baseline. G, Grip test to detect neuromuscular function. The Y-axis is the normalized moved distance baseline. The black arrow indicates drug infusion. H, Flow analysis of ATF3 expression of primary DRG cells at week 4 for NeuN+-ATF3+ double staining. ****P < 0.0001, ***P < 0.001, **P < 0.01, *P < 0.05 compared with baseline using a two-tailed Student's t-test. Each value is represented as mean ± SEM from at least seven mice in each group.

Fig 4: Effects of oxaliplatin-induced remodeling of calcium homeostasis on CTSS expression via IRF1. A, The effect of oxaliplatin on the nuclear localization of IRF1. Cells were treated with or without oxaliplatin (1 µM) for 24 h, and the cytosolic and nuclear fractions were then separated, as described in the Materials and Methods section. B, The effect of oxaliplatin treatment for 24 h on IRF1 binding to the CTSS promoter regions in SIM-A9 cells was analyzed using ChIP. C, The effect of oxaliplatin treatment for 24 h on the transcriptional activity of CTSS promoters was determined using the promoter assay. D, The effect of oxaliplatin treatment for 24 h on the transcription levels of CTSS genes in SIM-A9 was determined using RT-qPCR. E, The effect of oxaliplatin on the transcription levels analysis of CTSS genes in mouse primary DRG at week 4 was determined using RT-qPCR. F, The time-dependent mouse serum CTSS level (n = 10). G, Patients with colorectal cancer' serum CTSS level at week 48 compared to baseline (n = 23). ****P < 0.0001, Two-way ANOVA analysis. H, For the drug treatment, the cells were pretreated with DMSO or RJW-58 (5 µM). The treatment time for ND 7/23 and SIM-A9 was 24 h. The calcium influx from the ER and SOCE in CTSS overexpression (n = 50 for each condition) cell was assessed using single-cell intracellular Ca2+ measurement. I, CTSS was disassociated from STIM1 following RJW-58 treatment. ****P < 0.0001, ***P < 0.001, **P < 0.01, *P < 0.05. The statistical significance was determined using a two-tailed Student's t-test.