rabbit polyclonal antibodies against erα  (Proteintech)

Journal: Frontiers in Immunology

Article Title: SNRPB2 facilitates esophageal squamous cell carcinoma oncogenesis and progression via E2F4 stabilization

doi: 10.3389/fimmu.2025.1610721

Figure Lengend Snippet: SNRPB2 was highly expressed in ESCA which was associated with unfavorable clinical characteristics and poor prognosis. (a) Kaplan-Meier curves were used to analyze the influence of SNRPB2 mRNA on OS and DSS in ESCA patients. (b) Relative SNRPB2 mRNA expression in paired normal and tumor tissues from ESCA patients. (c) Representative IHC images of SNRPB2 protein expression in tumor and para-carcinoma tissues of ESCC. The upper panel shows low-grade ESCC and the lower panel high-grade ESCC, each with para-carcinoma tissue and tumor samples. Scale bars: 100 μm. (d) The expression levels of SNRPB2 protein in para-carcinoma tissues versus ESCC tissues. (e) Association of SNRPB2 protein expression with tumor grade and stage in ESCC. (f) The correlation between SNRPB2 protein expression and OS of ESCC. (g) Protein and relative mRNA levels of SNRPB2 in normal esophageal epithelial cells and ESCC cell lines were determined by western blot and q-PCR, with β-actin as the loading control. Data represent mean ± SD from three independent experiments. Statistical analysis was performed using Student’s t-test and two-sided t-test. (* P < 0.05; ** P < 0.01; *** P < 0.001).

Article Snippet: The in situ protein expression levels of SNRPB2 and E2F4 in paraffin-embedded ESCC tissue sections were assessed by immunohistochemistry using rabbit polyclonal antibodies against SNRPB2 (1:100, 21244-1-AP, Proteintech) and E2F4 (1:100, 10923-1-AP, Proteintech).

Techniques: Expressing, Western Blot, Control

Journal: Frontiers in Immunology

Article Title: SNRPB2 facilitates esophageal squamous cell carcinoma oncogenesis and progression via E2F4 stabilization

doi: 10.3389/fimmu.2025.1610721

Figure Lengend Snippet: SNRPB2 promoted proliferation, migration and invasion of ESCC cells. (a) Western blot analysis of SNRPB2 protein levels in TE-1 and KYSE-150 cells after transduction with two independent shRNAs targeting SNRPB2, with β-actin as the loading control. Bar graphs on the right show quantification of relative SNRPB2 protein levels normalized to β-actin. (b) The number of colony cells decreased significantly when cells were treated with SNRPB2 shRNAs. (c) The cell viability significantly decreased when cells were treated with SNRPB2 shRNAs. (d, e) TE-1 and KYSE-150 cells transfected with SNRPB2 shRNAs were evaluated by transwell migration assay, matrigel invasion assay, and wound healing experiment. All data were presented as the mean ± SDs (n = 3). Statistical analysis was performed using Student’s t-test. (** P < 0.01; *** P < 0.001).

Article Snippet: The in situ protein expression levels of SNRPB2 and E2F4 in paraffin-embedded ESCC tissue sections were assessed by immunohistochemistry using rabbit polyclonal antibodies against SNRPB2 (1:100, 21244-1-AP, Proteintech) and E2F4 (1:100, 10923-1-AP, Proteintech).

Techniques: Migration, Western Blot, Transduction, Control, Transfection, Transwell Migration Assay, Invasion Assay

Journal: Frontiers in Immunology

Article Title: SNRPB2 facilitates esophageal squamous cell carcinoma oncogenesis and progression via E2F4 stabilization

doi: 10.3389/fimmu.2025.1610721

Figure Lengend Snippet: SNRPB2 Expression Is Associated with Immune Infiltration and Immune Gene Signatures in ESCC. (a) Single-cell expression distribution of SNRPB2 across different cell types in ESCA, based on dataset GSE160269 . (b) Correlation between SNRPB2 expression and immune cell infiltration levels in ESCA using the TIMER database. (c) Correlation between SNRPB2 expression and representative immune-related genes from GEPIA2. (d) Relative mRNA expression levels of immune-related genes in TE-1 and KYSE-150 cells after SNRPB2 knockdown, analyzed by qPCR. Data represent mean ± SD from three independent experiments. Statistical analysis was performed using Student’s t-test. (* P < 0.05; ** P < 0.01; *** P < 0.001; ns, not significant).

Article Snippet: The in situ protein expression levels of SNRPB2 and E2F4 in paraffin-embedded ESCC tissue sections were assessed by immunohistochemistry using rabbit polyclonal antibodies against SNRPB2 (1:100, 21244-1-AP, Proteintech) and E2F4 (1:100, 10923-1-AP, Proteintech).

Techniques: Expressing, Knockdown

Journal: Frontiers in Immunology

Article Title: SNRPB2 facilitates esophageal squamous cell carcinoma oncogenesis and progression via E2F4 stabilization

doi: 10.3389/fimmu.2025.1610721

Figure Lengend Snippet: SNRPB2 inhibited Rb/E2F pathway in ESCA and interacted with E2F4 in ESCC cells. (a) Pathway enrichment: GSEA enrichment analyses of co‐expressed genes indicating an association of SNRPB2 with the top six signaling pathways in the TCGA-ESCA cohort. (b) GO functional enrichment analysis of SNRPB2 and its interactors were performed. (c) Correlation of the mRNA levels of SNRPB2 and E2F4 in TCGA-ESCA tumor samples. (d) The expression level of E2F4 mRNA in ESCA and normal esophageal tissue samples. (e) The Co-IP assay demonstrated the specific interactions between SNRPB2 and E2F4 in TE-1 and KYSE-150 cells. IgG was used as a negative control; whole cell lysates served as input. (f) Western blot analysis of E2F4 and SNRPB2 protein levels after treatment with MG132 or CQ in shCtrl or shSNRPB2 TE-1 cells. β-actin was used as the loading control. (g) The protein level of E2F4 after treated with CHX was measured by western blot in TE-1 and KYSE-150 cells, with β-actin as the internal control. Quantification of E2F4 band intensity normalized to β-actin is shown (right). (h) qPCR analysis of E2F4 target gene expression in TE-1 and KYSE-150 cells with or without SNRPB2 knockdown. β-actin was used for normalization. Data are represented as the mean ± SD (n = 3). Statistical analysis was performed using Student’s t-test. (* P < 0.05; ** P < 0.01; *** P < 0.001; ns, not significant).

Article Snippet: The in situ protein expression levels of SNRPB2 and E2F4 in paraffin-embedded ESCC tissue sections were assessed by immunohistochemistry using rabbit polyclonal antibodies against SNRPB2 (1:100, 21244-1-AP, Proteintech) and E2F4 (1:100, 10923-1-AP, Proteintech).

Techniques: Protein-Protein interactions, Functional Assay, Expressing, Co-Immunoprecipitation Assay, Negative Control, Western Blot, Control, Targeted Gene Expression, Knockdown

Journal: Frontiers in Immunology

Article Title: SNRPB2 facilitates esophageal squamous cell carcinoma oncogenesis and progression via E2F4 stabilization

doi: 10.3389/fimmu.2025.1610721

Figure Lengend Snippet: E2F4 overexpression rescued the effects of shSNRPB2 on proliferation, migration and invasion of ESCC cells in vitro and in vivo . (a, b) shCtrl+E2F4 group increased the proliferation and clone formation of ESCC compared with shCtrl+Vector group, and shSNRPB2+E2F4 group also increased the proliferation and clone formation of ESCC compared with shSNRPB2+Vector group. (c, d) shCtrl+E2F4 group promoted the migration and invasion of ESCC compared with shCtrl+Vector group, and shSNRPB2+E2F4 group accelerated the migration and invasion of ESCC compared with shSNRPB2+Vector group. (e) Five nude mice in each group were subcutaneously injected with TE-1 cells with shCtrl+Vector, shSNRPB2+Vector, shCtrl+E2F4, and shSNRPB2+E2F4. The mean tumor volume (cm 3 ) and weight (g) were measured three weeks later. (f) Representative images of SNRPB2 and E2F4 protein expression in ESCC specimens. Scale bars: upper panels, 500 µm; lower panels, 100 µm. A positive correlation between SNRPB2 and E2F4 expression was observed in 125 ESCC samples. All in vitro experiments were performed in triplicate (n = 3); β-actin was used as an internal control where applicable. In vivo tumor data represent n = 5 mice per group. Data are presented as mean ± SD. Statistical significance was determined using unpaired Student’s t-test or chi-square test. (** P < 0.01; *** P < 0.001).

Article Snippet: The in situ protein expression levels of SNRPB2 and E2F4 in paraffin-embedded ESCC tissue sections were assessed by immunohistochemistry using rabbit polyclonal antibodies against SNRPB2 (1:100, 21244-1-AP, Proteintech) and E2F4 (1:100, 10923-1-AP, Proteintech).

Techniques: Over Expression, Migration, In Vitro, In Vivo, Plasmid Preparation, Injection, Expressing, Control

Journal: RNA biology

Article Title: Estrogen receptor alpha (ERα) regulates PARN-mediated nuclear deadenylation and gene expression in breast cancer cells.

doi: 10.1080/15476286.2024.2413821

Figure Lengend Snippet: Figure 1. Estrogen receptor alpha (ERα) is an activator of PARN-mediated nuclear deadenylation in MCF7 (ERα+) cells. (A) nuclear extracts (NEs) for cells treated with different concentrations of 17β-estradiol (E2) for the indicated times were used in in vitro deadenylation assays with radiolabeled capped L3(A30) RNA substrate. Purified RNA was analysed by denaturing PAGE. Left panel: representative deadenylation reactions from three independent biological assays are shown. Positions of the polyadenylated RNA L3(A30) and the L3 deadenylated product are indicated. Right panel: bar graph of relative deadenylation (RD) is shown. (B–C) in vitro deadenylation assays using NEs from cells treated with (B) control (CTRL) or ERα siRNA for 24 h or (C) with increasing concentrations of fulvestrant for 2 h (FVT) were performed and analysed as in (A). (D) MCF7 cells were treated with either CTRL or PARN siRNA and subsequently treated with vehicle or E2. NEs were used for in vitro deadenylation as performed and analysed in (A). E) Cell-free deadenylation assays were performed in the presence of radiolabeled capped L3(A30) RNA substrates, limiting amount of his-PARN deadenylase and his-ERα and increasing amounts of GST-p53. Conditions for deadenylation assays were performed as in (A). F) NEs from untreated cells were used in endogenous reciprocal co-immunoprecipitation (e-ip) assays with polyclonal ERα, PARN, or p53 antibodies. NEs were treated with RNase A. Equivalent amounts of pellets (IP) and supernatants (SN) were resolved by SDS-PAGE, and proteins were detected by Western blot. Topo II was used as loading and IP specificity control. Ten percent of the NEs used in the e-ip assays are shown as input. All figures show representative deadenylation reactions and Western blot analyses from at least three independent biological assays analysed by triplicate (n = 3). Experiments with two groups were analysed using two-tailed unpaired Student’s t-test. The p-values are indicated as *(<0.01), **(<0.001) and ***(<0.0001).

Article Snippet: NEs were IPed with antibodies against ERα polyclonal (Cell Signaling, cat# 8644S), PARN polyclonal (Bethyl Laboratories, cat# A303-562A), or p53 polyclonal (Santa Cruz Biotechnology, cat# FL-393) using protein A-magnetic beads (Millipore, PureProteome cat# LSKMAGA10) per manufacturer’s instructions.

Techniques: In Vitro, Purification, Control, Immunoprecipitation, SDS Page, Western Blot, Two Tailed Test