Fig 1: Expression of HUWE1 in CD4+ T cells in peripheral blood from immune thrombocytopenic purpura patients. (A) Quantitative real-time PCR (qRT-PCR) and Western blot assays were performed to detect the mRNA and protein levels of HUWE1 in CD4+ T cells in the peripheral blood from healthy controls and immune thrombocytopenic purpura (ITP) patients; and the quantitative analysis of HUWE1 protein level (mean ± SEM, n = 5). (B) Correlation analysis of the mRNA level of HUWE1 and platelet counts in CD4+ T cells in the peripheral blood from ITP patients (r = -0.890, p < 0.01), n = 30. (C) Flow cytometry was applied to analyze the percentage of Treg cells in CD4+ T cells in peripheral blood from ITP patients and correlation analysis of the mRNA level of HUWE1 and the percentage of Treg cells in CD4+ T cells in peripheral blood from ITP patients (r = -0.858, p < 0.01), n = 30. (D) Western blot was performed to assess the Ets-1 protein level in the CD4+ T cells in the peripheral blood from ITP patients. The experiment was repeated three times. GAPDH is applied as the loading control. **p < 0.01 vs. control. ITP, immune thrombocytopenic purpura.
Fig 2: Differentiation and Y-chromosome detection in CPC-based cell sheets. (a, b) Differentiation of CPC in cell sheets toward vascular lineage. Representative images of immunofluorescence staining of the heart sections with antibodies against endothelial marker Ets-1 (a) and CD31 (b). Grafted cells were identified by CM-DIL (red fluorescence). Arrows indicate CM-DIL+ vessels. (c) Y-chromosome detection in CPC-derived endothelial cell. Representative images of immunofluorescent staining of heart sections with antibodies against endothelial marker CD31 and Y-chromosome. Arrows indicate Y-chromosome+ endothelial cells. (d, e) Differentiation of CPC in cell sheets toward cardiomyocyte lineage. Representative images of immunofluorescent staining of heart sections with antibodies against cardiomyocyte markers Nkx2.5 (d) and MHC (e) (ß-myosin heavy chain). Grafted cells were identified by CM-DIL (red fluorescence). Arrows indicate CM-DIL+ cardiomyocytes. (f) Y-chromosome detection in cardiomyocytes. Representative images of immunofluorescence staining of the heart sections with antibodies against cardiomyocyte marker MHC and Y-chromosome probe. Arrows indicate Y-chromosome+ cardiomyocytes.
Fig 3: Overexpression of SLC26A4-AS1 induces autophagy in PTC through promoting ETS1. A, Western blot analysis of autophagy related protein LC3-I, LC3-II, and p62 expression in oe-ITPR1-treated cells. B, Western blot analysis of autophagy related protein LC3-I, LC3-II, and p62 expression in oe-SLC26A4-AS1-treated cells upon Bafilomycin A1. C, Western blot analysis of Beclin-1 and Bcl2 expression upon oe-SLC26A4-AS1. * P < .05, **P < .01, and *** P < .001. Measurement data were expressed as mean ± standard deviation. The cancer tissues and adjacent normal tissues were compared by paired t test while analysis of the other two group was performed through unpaired t test. Analysis among multiple groups was conducted by ANOVA followed by Tukey's post hoc test
Fig 4: SLC26A4-AS1 represses tumour growth through up-regulation of ITPR1 by ETS1. A, Representative macroscopic images of mouse tumour upon treatment of oe-SLC26A4-AS1, oe-SLC26A4-AS1 + sh-ETS1 or controls. B, Quantification of tumour weight upon treatment of oe-SLC26A4-AS1, oe-SLC26A4-AS1 + sh-ETS1 or controls. C, RT-qPCR analysis of SLC26A4-AS1 and ETS1 expression in oe-SLC26A4-AS1- and sh-ETS1-treated mice and or controls. D, Immunohistochemistry of ITPR1 protein expression upon all treatments (×400). * P < .05 vs. oe-NC, ** P < .01, and *** P < .001. Measurement data were expressed as mean ± standard deviation. Analysis among multiple groups was conducted by ANOVA followed by Tukey's post hoc test. Data at different time points among groups were compared by repeated measures ANOVA, followed by Bonferroni
Fig 5: Regulation of HUWE1 on the differentiation and function of Treg cells through Ets-1. Naive CD4+ T cells were isolated from the peripheral blood of healthy controls and transfected with HUWE1 lentivirus and/or Ets-1 lentivirus for 48 h, and were cultured in Treg polarization conditions (cells in the presence of plate-bound anti-CD3, solid anti-CD28, TGF ß (5 ng/ml), and IL-2 (50 IU/ml) for 72 h. (A) Western blot was applied to measure the protein levels of HUWE1 and Ets-1. (B) qRT-PCR was performed to quantify the mRNA level of Foxp3 (mean ± SEM, n = 3). (C) Flow cytometry was conducted to assess the percentage of Treg cells in Naive CD4+ T cells (mean ± SEM, n = 8). (D) Treg cells from the peripheral blood of healthy controls were transfected with HUWE1 lentivirus and/or Ets-1 lentivirus for 48 h and then cultured with effector T cells in a ratio of 1:4. Immunosuppression assay was performed to analyze the inhibitory effect of Treg cells on the proliferation of effector T cells (mean ± SEM, n = 8). **p < 0.01 vs. Lenti or Lenti-HUWE1. The experiment was repeated three times.
Supplier Page from Abcam for Anti-ETS1 antibody