Fig 1: PRMT1 enhances chemoresistance of esophageal TICs.a, b Flow cytometric analysis showed that the percentage of OV6+ cells was significantly increased in LV-PRMT1 cells compared with LV-GFP cells. Data are shown as the mean ± SD, *P < 0.05, **P < 0.01. c, d Flow cytometric analysis revealed that the standard 4 days of chemotherapy with cis-platinum (2.5 g/ml) augmented the percentage of OV6+ subpopulation in ECA109 and TE1 cells. Data are presented as mean ± SD *P < 0.05, **P < 0.01. DDP, cis-platinum. e Cyclocystokinin-8 assay (CCK8) indicated that PRMT1 overexpression enhanced the cell viability of ECA109 and TE1 OV6+ cells in the presence of cis-platinum (0.6 g/ml). Data are shown as the mean ± SD, P < 0.01. f, g Increased ability of colony formation was also observed in LV-PRMT1 groups compared with LV-GFP groups after treated with cis-platinum (0.6 g/ml) for 14 days. Data are presented as the mean ± SD *P < 0.05, **P < 0.01. All experiments were performed in triplicates
Fig 2: Knockdown of PRMT1 represses the activation of IL‐6 trans‐signaling. (A) Western blot detection of PRMT1, p‐STAT3, and total STAT3 in mRTECs treated with IL‐6 or IL‐6 and sIL‐6R complex and the relative expression levels of p‐STAT3 and PRMT1. **P < 0.01, ***P < 0.001, and ****P < 0.0001 vs. NC. (B) Western blot assay of PRMT1, COX‐2, p‐STAT3, and total STAT3 and their relative expression in si‐PRMT1‐transfected mRTECs under L‐6 and sIL‐6R complex treatment. Data are expressed as the mean ± SD and analyzed using one‐way ANOVA with Tukey's post‐hoc test. All data were obtained from three independent experiments. *P < 0.05 and **P < 0.01 vs. si‐NC without IL‐6/sIL‐6R. PRMT1, protein arginine methyltransferase‐1; sIL‐6R, soluble IL‐6R.
Fig 3: FRAP analysis of PRMT1 kinetics in nucleus, cytoplasm, and PRMT1 cytoplasmic bodies. A) PRMT1 dynamics in untreated control U2OS cells. B) PRMT1 dynamics in untreated control HeLa cells. C) PRMT1 dynamics in U2OS cells treated with MC 1981 for 4 h. D) PRMT1 dynamics in U2OS cells treated with MC 2089 for 1-2 h. Asterisks mark statistically significant differences from values measured for PRMT1 dispersed in the cytoplasm (P≤0.05). Statistical analyses were performed using Student’s t-test on data normalized to 1; results are given as mean ± SEM.
Fig 4: PRMT1 expression is upregulated in TC cells. Western blotting was used to detect the expression of PRMT1 in human TC cell lines 8505C and CAL62, and in the human normal thyroid cell line Nthy-ori 3-1. **P<0.01 vs. Nthy-ori 3-1. PRMT1, protein arginine methyltransferase 1; TC, thyroid carcinoma.
Fig 5: PRMT1 knockdown suppresses cell metastasis and downregulates expression of ZEB1 and H4R3me2as in TC. (A) IF analyzed the contents of E-cadherin and vimentin in 8505C cells, magnification=400×. (B) Western blotting was used to assess the protein expression levels of E-cadherin, vimentin, H4R3me2as, and ZEB1 in 8505C cells. The 8505C cells were transfected with si-NC or si-PRMT1. *P<0.05, **P<0.01 compared with si-NC; ##P<0.01 compared with si-PRMT1; ^^P<0.01 compared with 10 µM AMI-1. Each experiment was conducted at least thrice. PRMT1, protein arginine methyltransferase 1; ZEB1, zinc finger e-box binding homeobox1, H4R3me2as, asymmetric demethylation of H4 at the third arginine residue; TC, thyroid carcinoma; si-NC, small interfering RNA negative control.
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