EZH2 Antibody - #AF5150

Fig. 1. EZH2 expression in PBMC and CD4+ T cells from RA and HC. (A) EZH2 mRNA expression in PBMC, CD4+ T cells, CD14+ monocytes and CD19+ B cells from HCs and RA patients (n = 22). (B) EZH2 protein expression in peripheral CD4+cells from HCs and RA patients (n = 5). (C) EZH2 protein expression in peripheral CD4+ CD45RA+ naïve T cells from HCs and RA patients (n = 20). (**: p < 0.01, ****: p < 0.0001).

Figure 4 m6A-associated pseudogene can regulate targeted immune-involved genes via miRNAs. (A) Sankey showing pseudogenes together with binding miRNAs and target genes with | R | ≥ 0.3 and P < 0.05 were used to construct the pseudogene-miRNA-target gene regulatory networks by subtypes of oncogene pseudogene PDIA3P1 and tumor-suppressor pseudogene RRN3P3. The column on the left represented pseudogenes, which are located at the cores of the networks. The column in the middle and the column on the right stand for binding miRNAs and target genes, respectively. (B-G) Experimental validation of PDIA3P1 affects the expression of AKT1 via miR-34a-5p in ARO and Tca8113 cell lines. (B) Relative gene expression of PDIA3P1 after PDIA3P1 knockdown using siRNA. (C) Relative gene expression of AKT1 after PDIA3P1 knockdown using siRNA. (D) Western blot comparing the protein levels of AKT1 in control and PDIA3P1 knockdown cells. (E) The relative expression of AKT1 at different time points after transcription inhibition in control and PDIA3P1 knockdown cells respectively. Error bars represent standard errors. (F) The relative expression of PDIA3P1 and AKT1 after adding control inhibitor versus miR-34a-5p inhibitor. (G) Western blot comparing the protein levels of AKT1 after adding control inhibitor versus miR-34a-5p inhibitor. (H-M) Experimental validation of RRN3P3 affects the expression of EZH2 via miR-26b-5p in ARO and Tca8113 cell lines. (H) Relative gene expression of RRN3P3 after RRN3P3 knockdown using siRNA. (I) Relative gene expression of EZH2 after RRN3P3 knockdown using siRNA. (J) The relative expression of EZH2 at different time points after transcription inhibition in control and RRN3P3 knockdown cells respectively. Error bars represent standard errors. (K) Western blot comparing the protein levels of EZH2 in control and RRN3P3 knockdown cells. (L) The relative expression of RRN3P3 and EZH2 after adding control inhibitor versus miR-26b-5p inhibitor. (M) Western blot comparing the protein levels of EZH2 after adding control inhibitor versus miR-26b-5p inhibitor. (N) UCSC genome browser tracks m6A-seq and m6A-LAIC-seq data indicating m6A peaks and m6A levels of oncogene pseudogene PDIA3P1 and tumor-suppressor pseudogene RRN3P3. Read-coverage tracks of input, m6A-negative, and m6A-positive fractions of m6A-LAIC-seq shown along with overlay tracks of m6A-seq (cyan for input and red for RIP; predicted m6A sites in m6A peaks are indicated by arrows). Read coverage (y-axis) of m6A negative and m6A positive are normalized as previously described 17 to reflect the calculated m6A levels (i.e., equal signals in m6A positive (eluate) versus m6A negative (supernatant) suggest m6A levels of 50%), while input and IP tracks of m6A-seq are shown for optimal viewing at the top panel. * P< 0.05; ** P< 0.01; *** P< 0.001 (two-tailed t-test). (O) The m6A methylation level of the pseudogenes at specific modification sites (Chr1:146650342, GG(m6A)CA on PDIA3P1; Chr16:22431201, GG(m6A)CG on RRN3P3) using SELECT in control and METTL3 knockdown ARO and Tca8113 cell.

Figure 3.| Pathological staining for evaluating the severity of AF in rats. (C) Immunohistochemistry assay was utilized to examine EZH2 expression in atrial tissues. Scale bar, 50 µm.

Figure 1. Expression levels of miR-101a-3p and EZH2 in atrial tissues of rats. The rats were anesthetized with an intraperitoneal injection of pentobarbital sodium (50 mg/kg), fixed, intubated and injected with LV-NC or LV-miR-101a-3p. After injection for 48 h, all rats were anesthetized. Then, 1 ml/kg acetylcholine-CaCl2 mixture via the tail vein was injected into rats daily for 7 days. At day 8, rats were anesthetized, injected with Ach-CaCl2 and sacrificed. The atrial tissues of heart were collected for following experiments. (A) Experimental protocol in rats. (B) miR-101a-3p expression was detected via reverse transcription-quantitative PCR. (C) Protein expression level of EZH2 was measured via western blotting. β-actin was used as an internal reference. The results are presented as the mean ± standard deviation (n=6). ##P<0.01 and ###P<0.001 vs. sham group; ***P<0.001 vs. AF + LV-NV group. LV-NC, lentivirus negative control; AF, atrial fibrillation; miR, microRNA; EZH2, enhancer of zeste 2 homolog 2; SD, Sprague-Dawley.

Figure 3 A: Showing invasive nests of basaloid cells in a case of basaloid SCC (H&E x4). B: Corresponding IHC in the same case showing strong nuclear positivity of EZH2 (Score 9) (IHC EZH2 x20). C: Showing papillae with fibrovascular core and lined by squamous epithelium in papillary SCC (H&E x10). D Corresponding IHC showing strong nuclear positivity of EZH2 (Score 9) (IHC EZH2, x20).