Fig 2: GR/miR-143-3p/JAG1/NOTCH2 regulates tumorigenesis of CRPC cells in vivo. A, Representative images of tumors formed in nude mice 40 days after injection with CRPC cells. B, The tumor growth curve. C. The final tumor size in nude mice. D, The expression of miR-143-3p in tumor tissues was determined by RT-qPCR. E, The expression of GR, Bcl-2, Cyclin D1, Cleaved-PARP, Cleaved-Caspase 3, JAG1, NOTCH2, E-cadherin and Vimentin normalized to GAPDH in tumor tissues was measured by Western blot analysis. In panel C, D, and E, p*< 0.05 compared with sh-NC group. All the measurement data were expressed as mean ± standard deviation. The paired t-test (n= 5) was used to compare the data of the two groups of paired designs, and the unpaired t-test was used to compare the data of the two groups of unpaired designs that followed normal distribution and homogeneity of variance. Data of different groups at different time points were compared by repeated measures ANOVA, and Bonferroni was used for post-test. N= 5.

Fig 3: Differential regulation of Notch signaling genes by CII and X-rays in human glioma cell lines: The mRNA expression of Notch receptors (relative to un-irradiated sample): Notch1 (N1), Notch2 (N2), and Notch3 (N3); Notch target genes: Hes1 and Hey1, observed at 24 h after irradiation were analyzed by qRT-PCR in LN229 and U251 monolayer cell culture irradiated with either (A,C) 2 Gy X-ray or carbon ion irradiation or; (B,D) 4 Gy X-ray or carbon ion irradiation. Each symbol represents an independent experiment performed in triplicate. Data are presented as mean ± SD. p values were determined by an unpaired two-tailed Student’s t-test, * p < 0.05, ** p < 0.01.

Fig 4: Notch receptors gene expression profiles in ex vivo pAEC from children with and without wheeze. Relative expression of NOTCH1–4 receptors (A–D) were measured in ex vivo pAEC from children with and without wheeze (n = 6) by real-time qPCR (refer to 2.5.4). (A) NOTCH1 was the most abundantly expressed Notch receptor gene in ex vivo pAEC from a paediatric cohort. However, NOTCH1 mRNA expression was not significantly different between pAEC from children without wheeze. (B) In contrast, mRNA expression of NOTCH2 was significantly lower in pAEC from children with wheeze compared to their non-wheeze counterparts. (C,D) Finally, no significant differences were observed between pAEC from children with and without wheeze in mRNA expression of NOTCH3 and NOTCH4, although the genes were expressed at different magnitudes. Each dot represents the mean of two replicates from a single ex vivo sample. Data are normalized to housekeeping gene, 18S and represented as median ± IQR. * Statistical significance relative to the non-wheeze cohort (p < 0.050; Mann–Whitney U-test).

Fig 5: GR was highly expressed in CRPC, while miR-143-3p was poorly expressed in CRPC. JAG1 and NOTCH2 were the target genes of miR-143-3p. GR could potentially elevate the expression of JAG1/NOTCH2 via inhibition of miR-143-3p. Silencing GR inhibited cell proliferation, migration and invasion through up-regulation of miR-143-3p, thereby suppressing EMT and progression of CRPC.