Journal: Frontiers in Immunology

Article Title: Case Report: Novel IRF2BP2 variant in a Japanese patient with impaired B-cell differentiation, Th1 polarization, and systemic immune dysregulation

doi: 10.3389/fimmu.2025.1662899

Figure Lengend Snippet: Identification of a novel IRF2BP2 mutation. (A) Sanger sequencing chromatograms of the IRF2BP2 gene showing a heterozygous de novo missense mutation (c.1663T>A; p. Cys555Ser) in the patient, which is absent in both parents. (B) Heatmap revealing the relative messenger RNA expression levels of IFI44L , LY6E , and MX1 in PBMCs from the patient and a healthy control, as measured using quantitative polymerase chain reaction. Expression levels are normalized to GAPDH and calculated using the comparative CT method. Red indicates higher expression, and blue indicates lower expression relative to the control. FC, fold change.

Article Snippet: All primers and probes were obtained from Applied Biosystems ( IFI44L [Hs00915292_m1], LY6E [Hs00158942_m1], MX1 [Hs00895608_m1], and GAPDH [Hs02758991_g1]).

Techniques: Mutagenesis, Sequencing, RNA Expression, Control, Real-time Polymerase Chain Reaction, Expressing

Journal: Cancer Immunology, Immunotherapy : CII

Article Title: Ly6E on tumor cells impairs anti-tumor T-cell responses: a novel mechanism of tumor-induced immune exclusion

doi: 10.1007/s00262-024-03851-x

Figure Lengend Snippet: Relationship between LY6E gene expression and clinical outcomes of patients with cancer. A The LY6E gene expression level was higher in 17 types of tumors compared to normal tissues. Each dot represents Ly6E gene expression in human normal (blue) and cancer tissues (red). B Kaplan–Meier curves of OS (overall survival) and DFS (disease-free survival) analyzed based on high- and low-Ly6E gene expression subgroups in 2376 cancer patient samples in 31 cancer types and 1137 cancer patient samples in 17 cancer types. C Kaplan–Meier survival curve of patients with high and low levels of LY6E gene expression level in tumor tissues before receiving anti-PD-1, anti-CTLA-4 or anti-PD-L1 antibodies. All data were obtained from http://kmplot.com/analysis . Overall survival probability was analyzed on the basis of cutoff values of the median ratio

Article Snippet: Ly6E expression levels on mouse cell lines were detected by flow cytometry using anti-Ly6E (#NBP1-52176, Novus Biologicals, USA).

Techniques: Expressing

Journal: Cancer Immunology, Immunotherapy : CII

Article Title: Ly6E on tumor cells impairs anti-tumor T-cell responses: a novel mechanism of tumor-induced immune exclusion

doi: 10.1007/s00262-024-03851-x

Figure Lengend Snippet: Ly6E expression level was negatively correlated with CD8 + T cell infiltration in human breast tumors. A The expression of Ly6E and CD8 in human breast tumor tissues was analyzed by immunohistochemical staining. “ + ”and “−” referring to positive and negative expression of Ly6E in tumor sites. B , C The expression of Ly6E on the cell surface of human breast tumor cell lines ( B ) and mouse tumor cell lines ( C ) was analyzed using anti-Ly6E antibodies by flow cytometry

Article Snippet: Ly6E expression levels on mouse cell lines were detected by flow cytometry using anti-Ly6E (#NBP1-52176, Novus Biologicals, USA).

Techniques: Expressing, Immunohistochemical staining, Staining, Flow Cytometry

Journal: Cancer Immunology, Immunotherapy : CII

Article Title: Ly6E on tumor cells impairs anti-tumor T-cell responses: a novel mechanism of tumor-induced immune exclusion

doi: 10.1007/s00262-024-03851-x

Figure Lengend Snippet: Ly6E promoted tumor growth and metastasis in subcutaneous mouse model and tail vein injection metastatic mouse model. A Preparation of Ly6E-knockout and Ly6E-overexpressing mouse tumor cell lines. Two Ly6E-knockdown mouse tumor cell lines (4T1-KO and GL261-KO) and one Ly6E-overexpressing mouse tumor cell line (MC38-OE) were selected, and Ly6E expression levels were confirmed by flow cytometry. B In vitro growth curve of Ly6E-knockdown and Ly6E-overexpressing cell lines (4T1-KO, GL261-KO or MC38-OE) compared with control cells (4T1-WT, GL261-WT or MC38-Mock). C – L Tumor growth and metastasis formation of the three paired tumor cell lines. 4T1-WT cells or 4T1-KO cells were injected into the breast fat pad of female BALB/c mice to establish an animal model of breast cancer metastasis. GL261 cells were injected into the right cerebral hemisphere of C57BL/6 mouse brain, and MC38 cells were intravenously injected into C57BL/6 mice. Tumor growth ( C ) and lung metastasis ( D , E ) of 4T1-KO and 4T1-WT ( n = 5) were examined. Tumor growth ( F ), survival rate ( G ) and tumor-free rate ( H ) of GL261-KO and GL261-WT-bearing mice were recorded every 3–7 days ( n = 5). Body weight ( I ), lung metastasis node ( J–L ) for mice injected MC38-OE or MC38-mock cells via the tail vein (n = 5). Lung specimens were dissected and photographed from tumor models ( D , J ) and performed H&E staining (scale bar, 100 μm for graph E, 250 μm for graph L). One representative example of two independent experiments was shown. ** p < 0.01; **** p < 0.0001

Article Snippet: Ly6E expression levels on mouse cell lines were detected by flow cytometry using anti-Ly6E (#NBP1-52176, Novus Biologicals, USA).

Techniques: Injection, Knock-Out, Knockdown, Expressing, Flow Cytometry, In Vitro, Control, Animal Model, Staining

Journal: Cancer Immunology, Immunotherapy : CII

Article Title: Ly6E on tumor cells impairs anti-tumor T-cell responses: a novel mechanism of tumor-induced immune exclusion

doi: 10.1007/s00262-024-03851-x

Figure Lengend Snippet: Differences in tumor growth rates between Ly6E-KO and Ly6E-WT cells were impaired in T cell-deficient mice. A , B GL261-WT cells or GL261-KO cells were intracranially injected into the right cerebral hemisphere of C57BL/6 mice and then treated with different antibodies (anti-CD8 Ab, anti-CD4 Ab or control IgG) at various time points (on day 1, 8 and 16). A In vivo bioluminescent imaging and computer-obtained luminescence (photons/second) of each tumor model mouse implanted with firefly luciferase transgenic GL261-WT cells or GL261-KO cells. B Tumor growth (left panel), tumor-free mice (middle panel) and survival of the mice (right panel) were recorded in different groups, n = 5. C , D The models of 4T1 breast cancer metastasis were established in female BALB/c mice and then treated with different antibodies (anti-CD8 Ab or control IgG). C Tumor diameters, the percentage of tumor-free mice and survival curve for mice were recorded in different groups, n = 5. D BALB/c mice were first injected with 4T1-KO cells in the right flank to generate primary tumors. Three months later, tumor-free mice and control mice were rechallenged and injected with 4T1-WT cells or 4T1-KO tumor cells, respectively. Tumor growth was monitored, n = 5. One representative example of two independent experiments is shown. * p < 0.05 and *** p < 0.001

Article Snippet: Ly6E expression levels on mouse cell lines were detected by flow cytometry using anti-Ly6E (#NBP1-52176, Novus Biologicals, USA).

Techniques: Injection, Control, In Vivo, Imaging, Luciferase, Transgenic Assay

Journal: Cancer Immunology, Immunotherapy : CII

Article Title: Ly6E on tumor cells impairs anti-tumor T-cell responses: a novel mechanism of tumor-induced immune exclusion

doi: 10.1007/s00262-024-03851-x

Figure Lengend Snippet: Proinflammatory chemokines released from Ly6E-positive tumor cells promoted macrophage migration and differentiation into immunosuppressive subsets. A – D Single-cell RNA sequencing (scRNA-seq) was performed on a total of 50,000 cells derived from 4T1-WT or 4T1-KO tumor tissues. BALB/c mice were subcutaneously injected with 4T1-WT cells or 4T1-KO cells, and CD45-positive cells were then sorted from tumor tissues on day 7. UMAP representation showing eight main cell clusters among CD45-positive cells between 4T1-WT and 4T1-KO tumor tissues ( A ). Box plots depicting proportion of B cells, dendritic cells, fibroblasts, mast cells, macrophage, MDSC, neutrophils or T cell pericytes from total cells between 4T1-WT and 4T1-KO tumor ( B ). UMAP representation showing 16 clusters of immune cell subsets clustered by the expression of marker genes ( C ). Dot plot depicting expression levels of specific genes across five clusters with marker-based lineage assignments ( D ). E A schematic transwell model presents the migration of mouse macrophage ANA-1 cells in response to cell culture media (CM) from tumor cells. F CM-induced migration of CFSE-labeled ANA-1 cells into the lower chamber were compared between MC38-OE and MC38-WT by microscopy (left graph) and quantified using Image J software (right graph). Scale bar = 100 μm. G Migration of ANA-1 cells to CM derived from B16-WT and B16-KO cells. H-I The expression of macrophage marker genes in BMDMs was induced by CM derived from MC38-OE/MC38-WT ( H ) or 4T1-WT and 4T1-KO cells ( I ) for 24 h, and mRNA levels were determined by RT-q PCR. BMDMs, bone marrow-derived macrophages; MRC1, macrophage mannose receptor 1 (also known as CD206), Inos, inducible nitric oxide synthase

Article Snippet: Ly6E expression levels on mouse cell lines were detected by flow cytometry using anti-Ly6E (#NBP1-52176, Novus Biologicals, USA).

Techniques: Migration, RNA Sequencing Assay, Derivative Assay, Injection, Expressing, Marker, Cell Culture, Labeling, Microscopy, Software

Journal: Cancer Immunology, Immunotherapy : CII

Article Title: Ly6E on tumor cells impairs anti-tumor T-cell responses: a novel mechanism of tumor-induced immune exclusion

doi: 10.1007/s00262-024-03851-x

Figure Lengend Snippet: Inferred intercellular communication networks in Ly6E-expressing tumors. Ly6E-expressing tumor cells release the proinflammatory chemokines to recruit monocyte into the TME and polarize them to M2-like macrophages. Immunoregulatory factors like Arg-1 are then released from M2-like macrophages, facilitating Treg infiltration. These immunosuppressive cells in the tumor vicinity exclude CD8 + T cells from tumor islet. Ly6E-knockout tumor cells release anti-inflammatory cytokines like IL-12a, which promote M1-like macrophage polarization and enhanced the infiltration of CD8 + T cells into tumor islet

Article Snippet: Ly6E expression levels on mouse cell lines were detected by flow cytometry using anti-Ly6E (#NBP1-52176, Novus Biologicals, USA).

Techniques: Expressing, Knock-Out

Journal: Scientific Reports

Article Title: Lymphocyte antigen 6 family member E suppresses apoptosis and promotes pancreatic cancer growth and migration via Wnt/β-catenin pathway activation

doi: 10.1038/s41598-024-70764-1

Figure Lengend Snippet: LY6E expression and relationship with clinical characteristics in PC. ( A ) LY6E expression between PC tissues and normal tissues across TCGA + GTEx, GSE16515, GSE32676, GSE15471, and GSE55463 datasets. ( B ) LY6E expression difference in PC with different clinical subgroups. ( C ) Clinical characteristic difference in LY6E low-expression (C1) and high-expression (C2) subgroups. ( D ) Correlation between LY6E expression, age, clinical stage, tumor grade, and survival status. *, p < 0.05.

Article Snippet: The following primary Abs were used: LY6E rabbit polyclonal Ab (ab201098, 1:1000, Abcam), β-catenin rabbit monoclonal Ab (ab32572, 1:1000, Abcam), GSK3β rabbit monoclonal Ab (ab32391, 1:1000, Abcam), p-GSK3β rabbit monoclonal Ab (ab75814, 1:1000, Abcam), E-cadherin rabbit monoclonal Ab (ab40772, 1:1000, Abcam), N-cadherin rabbit monoclonal Ab (ab76011, 1:1000, Abcam), Vimentin rabbit monoclonal Ab (ab92547, 1:1000, Abcam), Bcl-2 rabbit monoclonal Ab (ab32124, 1:1000, Abcam), Bax rabbit monoclonal Ab (ab32503, 1:1000, Abcam), Cleaved caspase3 rabbit monoclonal Ab (ab32042, 1:1000, Abcam) and GAPDH mouse monoclonal Ab (ab8245, 1:1000, Abcam).

Techniques: Expressing

Journal: Scientific Reports

Article Title: Lymphocyte antigen 6 family member E suppresses apoptosis and promotes pancreatic cancer growth and migration via Wnt/β-catenin pathway activation

doi: 10.1038/s41598-024-70764-1

Figure Lengend Snippet: Correlation of prognostic ability of LY6E expression. ( A ) Distribution, survival status plot, survival time of LY6E expression in TCGA-PAAD project. Red dots and blue dots represent patients with high and low expression of LY6E, respectively. ( B ) Survival analysis of LY6E low-expression and high-expression PC in TCGA-PAAD project. ( C ) ROC curves analyses between LY6E low-expression and high-expression PC in TCGA-PAAD project. ( D ) Univariate and multivariate Cox regression analyses in TCGA-PAAD project. ( E ) A nomogram of LY6E expression and age for PC. ( F ) A calibration plot for assessing the predictive ability of the nomogram at 1- and 3-year.

Article Snippet: The following primary Abs were used: LY6E rabbit polyclonal Ab (ab201098, 1:1000, Abcam), β-catenin rabbit monoclonal Ab (ab32572, 1:1000, Abcam), GSK3β rabbit monoclonal Ab (ab32391, 1:1000, Abcam), p-GSK3β rabbit monoclonal Ab (ab75814, 1:1000, Abcam), E-cadherin rabbit monoclonal Ab (ab40772, 1:1000, Abcam), N-cadherin rabbit monoclonal Ab (ab76011, 1:1000, Abcam), Vimentin rabbit monoclonal Ab (ab92547, 1:1000, Abcam), Bcl-2 rabbit monoclonal Ab (ab32124, 1:1000, Abcam), Bax rabbit monoclonal Ab (ab32503, 1:1000, Abcam), Cleaved caspase3 rabbit monoclonal Ab (ab32042, 1:1000, Abcam) and GAPDH mouse monoclonal Ab (ab8245, 1:1000, Abcam).

Techniques: Expressing

Journal: Scientific Reports

Article Title: Lymphocyte antigen 6 family member E suppresses apoptosis and promotes pancreatic cancer growth and migration via Wnt/β-catenin pathway activation

doi: 10.1038/s41598-024-70764-1

Figure Lengend Snippet: Relationship of LY6E and immunity. ( A ) Correlation coefficient value between LY6E expression and immune cell infiltration. ( B ) Immune cell infiltration difference between LY6E low-expression and high-expression PC. ( C , D , E , F , G , H , I , J , and K ) Relationship between LY6E expression and memory B cells, activated dendritic cells, macrophage M0, activated NK cells, plasma cells, CD8 + T cells, activated memory CD4 + T cells, naive B cells, and tumor mutation burden. ( L ) Correlation between LY6E and other immune-related genes. Red indicates positive correlation, while blue stands for negative correlation. The darker the color, the stronger the correlation coefficient. *, p < 0.05. **, p < 0.01. ***, p < 0.001.

Article Snippet: The following primary Abs were used: LY6E rabbit polyclonal Ab (ab201098, 1:1000, Abcam), β-catenin rabbit monoclonal Ab (ab32572, 1:1000, Abcam), GSK3β rabbit monoclonal Ab (ab32391, 1:1000, Abcam), p-GSK3β rabbit monoclonal Ab (ab75814, 1:1000, Abcam), E-cadherin rabbit monoclonal Ab (ab40772, 1:1000, Abcam), N-cadherin rabbit monoclonal Ab (ab76011, 1:1000, Abcam), Vimentin rabbit monoclonal Ab (ab92547, 1:1000, Abcam), Bcl-2 rabbit monoclonal Ab (ab32124, 1:1000, Abcam), Bax rabbit monoclonal Ab (ab32503, 1:1000, Abcam), Cleaved caspase3 rabbit monoclonal Ab (ab32042, 1:1000, Abcam) and GAPDH mouse monoclonal Ab (ab8245, 1:1000, Abcam).

Techniques: Expressing, Mutagenesis

Journal: Scientific Reports

Article Title: Lymphocyte antigen 6 family member E suppresses apoptosis and promotes pancreatic cancer growth and migration via Wnt/β-catenin pathway activation

doi: 10.1038/s41598-024-70764-1

Figure Lengend Snippet: LY6E is highly expressed in PC and knockdown of LY6E impairs PC cell growth in vitro. ( A ) IHC staining of LY6E expression in the tumor and normal tissues of PC patients. Scale bar = 100 μm. ( B ) Western blotting analysis of LY6E protein expression in tumor tissues and normal tissues of PC patients. GAPDH was used as a control. ( C ) Western blotting analysis of LY6E protein expression in a human pancreatic cell line (HPNE) and PC cell lines (PANC-1, BxPC-3, and Patu8988). GAPDH was used as the control. ( D ) qRT-PCR analysis of LY6E mRNA transcription in BxPC-3 and Patu8988 cells transfected with lenti-shLY6E-1 and lenti-shLY6E-2. GAPDH was used as the control. ( E ) Western blotting analysis of LY6E in BxPC-3 and Patu8988 cells transfected with lenti-shLY6E-1 and lenti-shLY6E-2. GAPDH was used as the control. ( F ) Cell viability at 0, 24, 48 and 72 h were detected after cell was seeded using the CCK-8 assay. OD450 nm absorbance was measured. *** represents the lenti-shLY6E-1 treatment group compared to NC, with p < 0.001. ### represents the lenti-shLY6E-2 treatment group compared to NC, with p < 0.001. ( G ) Colony formation analysis of BxPC-3 and Patu8988 cells that were transfected with lenti-shLY6E-1 and lenti-shLY6E-2 (14 d after cell seeding). The number of colonies in each well was counted. ( H ) IFC staining of Ki67 in BxPC-3 and Patu8988 cells transfected with lenti-shLY6E-1 and lenti-shLY6E-2.*, p < 0.05.

Article Snippet: The following primary Abs were used: LY6E rabbit polyclonal Ab (ab201098, 1:1000, Abcam), β-catenin rabbit monoclonal Ab (ab32572, 1:1000, Abcam), GSK3β rabbit monoclonal Ab (ab32391, 1:1000, Abcam), p-GSK3β rabbit monoclonal Ab (ab75814, 1:1000, Abcam), E-cadherin rabbit monoclonal Ab (ab40772, 1:1000, Abcam), N-cadherin rabbit monoclonal Ab (ab76011, 1:1000, Abcam), Vimentin rabbit monoclonal Ab (ab92547, 1:1000, Abcam), Bcl-2 rabbit monoclonal Ab (ab32124, 1:1000, Abcam), Bax rabbit monoclonal Ab (ab32503, 1:1000, Abcam), Cleaved caspase3 rabbit monoclonal Ab (ab32042, 1:1000, Abcam) and GAPDH mouse monoclonal Ab (ab8245, 1:1000, Abcam).

Techniques: Knockdown, In Vitro, Immunohistochemistry, Expressing, Western Blot, Control, Quantitative RT-PCR, Transfection, CCK-8 Assay, Staining

Journal: Scientific Reports

Article Title: Lymphocyte antigen 6 family member E suppresses apoptosis and promotes pancreatic cancer growth and migration via Wnt/β-catenin pathway activation

doi: 10.1038/s41598-024-70764-1

Figure Lengend Snippet: Knockdown of LY6E promotes apoptosis by regulating cleaved caspase 3, Bcl-2, and Bax protein expression in PC. ( A ) Cell apoptosis was detected in BxPC-3 and Patu8988 cells that were transfected with lenti-shLY6E-1 and lentishLY6E-2. Flow cytometry was used to determine the percentage of cells that were apoptotic. ( B ) Western blotting analysis of cleaved caspase 3, Bcl-2, and Bax protein expression in BxPC-3 and Patu8988 cells that were transfected with lenti-shLY6E-1 and lentishLY6E-2. GAPDH was used as the control. ( C ) IFC staining of cleaved caspase 3 in BxPC-3 and Patu8988 cells transfected with lenti-shLY6E-1 and lenti-shLY6E-2. *, p < 0.05.

Article Snippet: The following primary Abs were used: LY6E rabbit polyclonal Ab (ab201098, 1:1000, Abcam), β-catenin rabbit monoclonal Ab (ab32572, 1:1000, Abcam), GSK3β rabbit monoclonal Ab (ab32391, 1:1000, Abcam), p-GSK3β rabbit monoclonal Ab (ab75814, 1:1000, Abcam), E-cadherin rabbit monoclonal Ab (ab40772, 1:1000, Abcam), N-cadherin rabbit monoclonal Ab (ab76011, 1:1000, Abcam), Vimentin rabbit monoclonal Ab (ab92547, 1:1000, Abcam), Bcl-2 rabbit monoclonal Ab (ab32124, 1:1000, Abcam), Bax rabbit monoclonal Ab (ab32503, 1:1000, Abcam), Cleaved caspase3 rabbit monoclonal Ab (ab32042, 1:1000, Abcam) and GAPDH mouse monoclonal Ab (ab8245, 1:1000, Abcam).

Techniques: Knockdown, Expressing, Transfection, Flow Cytometry, Western Blot, Control, Staining

Journal: Scientific Reports

Article Title: Lymphocyte antigen 6 family member E suppresses apoptosis and promotes pancreatic cancer growth and migration via Wnt/β-catenin pathway activation

doi: 10.1038/s41598-024-70764-1

Figure Lengend Snippet: Knockdown of LY6E suppresses migration and invasion of PC by regulating vimentin, E-Cadherin, and N-Cadherin protein expression. ( A ) Wound healing assay of BxPC-3 and Patu8988 cells that were transfected with lenti-shLY6E-1 and lentishLY6E-2. ( B ) Transwell assay of BxPC-3 and Patu8988 cells that were transfected with lenti-shLY6E-1 and lentishLY6E-2. ( C ) Western blotting analysis of certain migration-related proteins including vimentin, E-Cadherin, and N-Cadherin in BxPC-3 and Patu8988 cells that were transfected with lenti-shLY6E-1 and lentishLY6E-2. GAPDH was used as the control. ( D ) IFC staining of vimentin in BxPC-3 and Patu8988 cells transfected with lenti-shLY6E-1 and lenti-shLY6E-2. *, p < 0.05.

Article Snippet: The following primary Abs were used: LY6E rabbit polyclonal Ab (ab201098, 1:1000, Abcam), β-catenin rabbit monoclonal Ab (ab32572, 1:1000, Abcam), GSK3β rabbit monoclonal Ab (ab32391, 1:1000, Abcam), p-GSK3β rabbit monoclonal Ab (ab75814, 1:1000, Abcam), E-cadherin rabbit monoclonal Ab (ab40772, 1:1000, Abcam), N-cadherin rabbit monoclonal Ab (ab76011, 1:1000, Abcam), Vimentin rabbit monoclonal Ab (ab92547, 1:1000, Abcam), Bcl-2 rabbit monoclonal Ab (ab32124, 1:1000, Abcam), Bax rabbit monoclonal Ab (ab32503, 1:1000, Abcam), Cleaved caspase3 rabbit monoclonal Ab (ab32042, 1:1000, Abcam) and GAPDH mouse monoclonal Ab (ab8245, 1:1000, Abcam).

Techniques: Knockdown, Migration, Expressing, Wound Healing Assay, Transfection, Transwell Assay, Western Blot, Control, Staining

Journal: Scientific Reports

Article Title: Lymphocyte antigen 6 family member E suppresses apoptosis and promotes pancreatic cancer growth and migration via Wnt/β-catenin pathway activation

doi: 10.1038/s41598-024-70764-1

Figure Lengend Snippet: Overexpression of LY6E suppresses apoptosis, enhances proliferation and migration of PC cells. ( A ) CCK8 assay indicated that overexpression of LY6E augmented PC proliferation. ( B ) Clone formation assay indicated more colonies in LY6E overexpression in PC than in control. (C) The effect of LY6E overexpression on PC apoptosis was analyzed by flow cytometry. ( D ) Transwell showed that LY6E increased the number of migrating PC. *, p < 0.05. **, p < 0.01. ***, p < 0.001.

Article Snippet: The following primary Abs were used: LY6E rabbit polyclonal Ab (ab201098, 1:1000, Abcam), β-catenin rabbit monoclonal Ab (ab32572, 1:1000, Abcam), GSK3β rabbit monoclonal Ab (ab32391, 1:1000, Abcam), p-GSK3β rabbit monoclonal Ab (ab75814, 1:1000, Abcam), E-cadherin rabbit monoclonal Ab (ab40772, 1:1000, Abcam), N-cadherin rabbit monoclonal Ab (ab76011, 1:1000, Abcam), Vimentin rabbit monoclonal Ab (ab92547, 1:1000, Abcam), Bcl-2 rabbit monoclonal Ab (ab32124, 1:1000, Abcam), Bax rabbit monoclonal Ab (ab32503, 1:1000, Abcam), Cleaved caspase3 rabbit monoclonal Ab (ab32042, 1:1000, Abcam) and GAPDH mouse monoclonal Ab (ab8245, 1:1000, Abcam).

Techniques: Over Expression, Migration, CCK-8 Assay, Tube Formation Assay, Control, Flow Cytometry

Journal: Scientific Reports

Article Title: Lymphocyte antigen 6 family member E suppresses apoptosis and promotes pancreatic cancer growth and migration via Wnt/β-catenin pathway activation

doi: 10.1038/s41598-024-70764-1

Figure Lengend Snippet: Knockdown of LY6E down-regulated the Wnt/beta-catenin signaling pathway. ( A ) Gene expression profile of LY6E low-expression and high-expression PC in TCGA-PAAD. G1 and G2 represent groups with high and low expression of LY6E mRNA. Columns and rows denote patients and genes. ( B ) Up-regulated and down-regulated genes in LY6E low-expression and high-expression PC in TCGA-PAAD. Red dots and blue dots indicate upregulated and downregulated genes. The x-axis represents fold change, while the y-axis reflects statistical significance. ( C ) GO/KEGG analysis of up-regulated and down-regulated genes in LY6E low-expression and high-expression PC in TCGA-PAAD. ( D ) Correlation analysis of LY6E expression and CTNNB1 expression. Points represent patients, with the horizontal and vertical axes displaying the expression levels of LY6E and CTNNB1, respectively. ( E ) Western blotting analysis of β-catenin, GSK3β, and p-GSK3β in BxPC-3 and Patu8988 cells that were transfected with lenti-shLY6E-1 and lentishLY6E-2. GAPDH was used as the control. ( F ) IFC staining of β-catenin in BxPC-3 and Patu8988 cells transfected with lenti-shLY6E-1 and lenti-shLY6E-2.

Article Snippet: The following primary Abs were used: LY6E rabbit polyclonal Ab (ab201098, 1:1000, Abcam), β-catenin rabbit monoclonal Ab (ab32572, 1:1000, Abcam), GSK3β rabbit monoclonal Ab (ab32391, 1:1000, Abcam), p-GSK3β rabbit monoclonal Ab (ab75814, 1:1000, Abcam), E-cadherin rabbit monoclonal Ab (ab40772, 1:1000, Abcam), N-cadherin rabbit monoclonal Ab (ab76011, 1:1000, Abcam), Vimentin rabbit monoclonal Ab (ab92547, 1:1000, Abcam), Bcl-2 rabbit monoclonal Ab (ab32124, 1:1000, Abcam), Bax rabbit monoclonal Ab (ab32503, 1:1000, Abcam), Cleaved caspase3 rabbit monoclonal Ab (ab32042, 1:1000, Abcam) and GAPDH mouse monoclonal Ab (ab8245, 1:1000, Abcam).

Techniques: Knockdown, Expressing, Western Blot, Transfection, Control, Staining

Journal: Scientific Reports

Article Title: Lymphocyte antigen 6 family member E suppresses apoptosis and promotes pancreatic cancer growth and migration via Wnt/β-catenin pathway activation

doi: 10.1038/s41598-024-70764-1

Figure Lengend Snippet: Knockdown of LY6E inhibits pancreatic tumor growth in vivo. ( A , B , and C ) Tumor image, end-stage tumor weight, and tumor volume ( D ) Western blotting analysis of Vimentin, E-Cadherin, and N-Cadherin protein expression in tumor tissues. GAPDH was used as the control. ( E ) HE staining and IHC staining of Ki67, β-catenin, and Cleaved caspase 3 in tumor tissues. *, p < 0.05.

Article Snippet: The following primary Abs were used: LY6E rabbit polyclonal Ab (ab201098, 1:1000, Abcam), β-catenin rabbit monoclonal Ab (ab32572, 1:1000, Abcam), GSK3β rabbit monoclonal Ab (ab32391, 1:1000, Abcam), p-GSK3β rabbit monoclonal Ab (ab75814, 1:1000, Abcam), E-cadherin rabbit monoclonal Ab (ab40772, 1:1000, Abcam), N-cadherin rabbit monoclonal Ab (ab76011, 1:1000, Abcam), Vimentin rabbit monoclonal Ab (ab92547, 1:1000, Abcam), Bcl-2 rabbit monoclonal Ab (ab32124, 1:1000, Abcam), Bax rabbit monoclonal Ab (ab32503, 1:1000, Abcam), Cleaved caspase3 rabbit monoclonal Ab (ab32042, 1:1000, Abcam) and GAPDH mouse monoclonal Ab (ab8245, 1:1000, Abcam).

Techniques: Knockdown, In Vivo, Western Blot, Expressing, Control, Staining, Immunohistochemistry

Journal: Scientific Reports

Article Title: Lymphocyte antigen 6 family member E suppresses apoptosis and promotes pancreatic cancer growth and migration via Wnt/β-catenin pathway activation

doi: 10.1038/s41598-024-70764-1

Figure Lengend Snippet: The rescue experiment verifies that LY6E promotes PC progression through Wnt/beta-catenin signaling pathway. ( A ) Transwell assay confirmed the restoration of invasion of BxPC-3 and Patu8988 cells transfected with lenti-shLY6E after being treated by LiCl. ( B and C ) Western blotting analysis of Vimentin, E-Cadherin, N-Cadherin, β-catenin, GSK3β, and p-GSK3β protein expression in BxPC-3 and Patu8988 cells transfected with lenti-shLY6E after treated by LiCl. GAPDH was used as the control. *, p < 0.05.

Article Snippet: The following primary Abs were used: LY6E rabbit polyclonal Ab (ab201098, 1:1000, Abcam), β-catenin rabbit monoclonal Ab (ab32572, 1:1000, Abcam), GSK3β rabbit monoclonal Ab (ab32391, 1:1000, Abcam), p-GSK3β rabbit monoclonal Ab (ab75814, 1:1000, Abcam), E-cadherin rabbit monoclonal Ab (ab40772, 1:1000, Abcam), N-cadherin rabbit monoclonal Ab (ab76011, 1:1000, Abcam), Vimentin rabbit monoclonal Ab (ab92547, 1:1000, Abcam), Bcl-2 rabbit monoclonal Ab (ab32124, 1:1000, Abcam), Bax rabbit monoclonal Ab (ab32503, 1:1000, Abcam), Cleaved caspase3 rabbit monoclonal Ab (ab32042, 1:1000, Abcam) and GAPDH mouse monoclonal Ab (ab8245, 1:1000, Abcam).

Techniques: Transwell Assay, Transfection, Western Blot, Expressing, Control