HES1 Antibody - #AF7575

Fig. 3. ALYREF promotes NPC metastasis by activating NOTCH1. A GSEA results of SUNE1 and SUNE2 cells with ALYREF knockdown (siALYREF) or siRNA control (siNC). B The relative Flag-RIP enrichment ratio of NOTCH1 mRNA in HONE1 cells with ALYREF overexpression or vector control. ACTB was used as the negative control. Fold enrichment was determined by calculating the 2-ΔCt of each RIP sample relative to the corresponding input sample. The mRNA and protein levels of NOTCH1 in NPC cells with ALYREF overexpression (C) and ALYREF knockdown (D) were determined using qRT-PCR and western blotting, respectively. E The mRNA and protein levels of downstream targets (HES1, HEY1) of the Notch signaling pathway in SUNE1 cells with ALYREF knockdown or the siNC control were determined by qRT-PCR and western blotting, respectively. F Representative immunohistochemical images of lung tissues isolated from mice injected with vector control HONE1 cells (left) or ALYREF-overexpressing cells (right) via the tail vein, confirming the high expression of NOTCH1 in the ALYREF overexpression group. G The protein levels of NOTCH1 in lung metastatic nodules with ALYREF overexpression or vector control in nude mice (n = 5 per group) were determined by western blot analysis. H Inhibition of the NOTCH1 pathway abrogated the ALYREF-mediated upregulation of NPC cell invasion and metastasis. HONE1-overexpressing and corresponding vector control cells were treated with 50 mg/L LY3039478 for 24 h. Scale bar: 100 μm. *P 

FIGURE 2 20 ng/mL IL-20 treated the pre-osteoclasts for 48 h meanwhile control groups were treated with 0 ng/mL IL-20. Detected mRNA and analysed by GO Analysis and Pathway Analysis. (A) Bubble chart analysis based on KEGG databases. (B) Gene Set Enrichment Analysis of osteoclast differentiation with control groups and IL-20 treat groups. (C) Gene heatmap showed four classical gene were enriched to Notch signal pathway (Red – up regulated and green – down regulated). (D-E) The expression of Notch1 and Notch2 mRNA with different concentration of IL-20 stimulation. Analysed by one-way ANOVA with Tukey's post hoc analysis with three independent experiments. *P < .05 vs the control group; ns, not significant. (F) mRNA expression of Notch1, Notch2, Hes1 and Hey1 at 20 ng/mL IL-20 stimulation using by qRT-PCR and analysed by Student's t test. Bars represent mean ± SEM from three independent experiments. *P < .05 vs the control group; ns, not significant. (G-H) Western blotting data and analysis of Notch signal pathway. Student's t test was used to detect difference. Bars represent mean ± SEM from three independent experiments. *P < .05 vs the control group; ns, not significant

Figure 9. - Protein levels of (A) hairy and enhancer of split 1, (B) PTEN, (C-1) p-AKT (Thr308), (C-2) AKT, (C-3) p-AKT (Ser473), (D-1) mTORC1, (D-2) p-mTOR (Ser2448), (E) IL-17A in mouse splenic mononuclear cells of control group, 0 µM LY294002 group, 10 µM LY294002 group and 50 µM LY294002 group. **P