Journal: Cell Death & Disease

Article Title: Atypical cadherin CELSR2 acts as a therapeutic target for glioma through WNT3A/β-catenin signaling

doi: 10.1038/s41419-025-08116-8

Figure Lengend Snippet: A Analysis of TCGA data reveals significantly elevated CELSR2 mRNA levels in primary and recurrent glioma tissues compared to normal tissues ( n = 1081 for normal; n = 662 for primary; n = 277 for recurrent). B Analysis of TCGA data demonstrates a significant upregulation of CELSR2 mRNA in low-grade glioma (LGG) and high-grade glioma (HGG) tissues relative to normal brain tissues ( n = 1081 for normal; n = 423 for LGG; n = 502 for HGG). C Analysis of TCGA data indicates that CELSR2 mRNA expression is significantly increased in Grade 2 and Grade 3 gliomas compared to normal brain tissues ( n = 1081 for normal; n = 115 for Grade 1; n = 289 for Grade 2; n = 254 for Grade 3; n = 248 for Grade 4). D RT-qPCR analysis shows elevated CELSR2 expression in clinical glioma tissues and glioma cell lines ( n = 3). N, para-tumoral normal tissue; T, tumor; T1: Grade 2; T2, T3: Grade 3; CP-H122, normal astrocyte line. E Kaplan–Meier survival curves illustrate reduced overall survival in patients with primary glioma (all WHO grades combined, n = 202). F Kaplan–Meier survival curves demonstrate decreased overall survival in patients with recurrent glioma (all WHO grades combined, n = 127 or 126). G Kaplan–Meier survival analysis reveals poorer prognosis in Grade 2 glioma patients (all WHO grades combined, n = 86). H Kaplan–Meier survival curve of grade 3 glioma patients (All WHO grade survival, n = 124). I Immunohistochemical staining showing CELSR2 levels in Grade 1-Grade 4 glioma tissues ( n = 3). Scale bar is 50 μm. Data are represented as mean ± SEM. ** P < 0.01, **** P < 0.0001, one-way ANOVA analysis of variance with Tukey’s multiple comparison in ( A – C ).

Article Snippet: The sections were incubated with antibody against CELSR2 (1:200, NOVUS, NLS1943) overnight at 4 °C, followed by incubation with the secondary antibody for 1 h.

Techniques: Expressing, Quantitative RT-PCR, Immunohistochemical staining, Staining, Comparison

Journal: Cell Death & Disease

Article Title: Atypical cadherin CELSR2 acts as a therapeutic target for glioma through WNT3A/β-catenin signaling

doi: 10.1038/s41419-025-08116-8

Figure Lengend Snippet: A Anti-GFAP (green) and anti-CELSR2 immunofluorescence staining showed the expression of CELSR2 in CP-H122 cells, U87 MG cells transfected with the vector (pLVX-shRNA1-ZsGreen-puro, control) or CELSR2 -shRNA ( CELSR2 KD), and the levels of CELSR2 mRNA were further assessed by RT-qPCR, showing a downregulation in U87 MG cells transfected with CELSR2 -shRNA. B – E EDU staining ( B , C ) and CCK-8 assay ( D , E ) showed a significant decrease in the number of proliferating cells in the CELSR2-KD group. F , G Two-dimensional culture assay showed reduced colonies in the CELSR2-KD group. H Using annexin V and propidium iodide (PI) staining, FACS analysis identified a comparable percentage of apoptotic cells in the CELSR2-KD group and the control group. I FACS analysis of PI-stained cells showed a significant increase in the G0/G1 ratio and a significant decrease in the S phase in the CELSR2-KD group. Scale bar is 100 μm in ( A , D ) and 200 μm in ( B ). Data are represented as mean ± SEM. * P < 0.05, ** P < 0.01, Two-way repeated measures ANOVA with Bonferroni’s post hoc correction in ( E ) and Student’s t -test in others, n = 3 in ( A – F ); n = 10 in ( H ); n = 5 in ( I ).

Article Snippet: The sections were incubated with antibody against CELSR2 (1:200, NOVUS, NLS1943) overnight at 4 °C, followed by incubation with the secondary antibody for 1 h.

Techniques: Immunofluorescence, Staining, Expressing, Transfection, Plasmid Preparation, Control, shRNA, Quantitative RT-PCR, CCK-8 Assay

Journal: Cell Death & Disease

Article Title: Atypical cadherin CELSR2 acts as a therapeutic target for glioma through WNT3A/β-catenin signaling

doi: 10.1038/s41419-025-08116-8

Figure Lengend Snippet: A – E DIA proteomics of cultured U87 MG cells transfected without (control) or with CELSR2 -shRNA ( CELSR2 KD) revealed 84 upregulated DEPs and 42 downregulated DEPs in the CELSR2-KD group compared to the control group ( A ; FDR < 0.05, |FoldChange| > 1.5), clustering in cancer pathways and the Wnt signaling pathway using KEGG pathway enrichment analysis ( B ). Downregulation of the cell cycle was shown by GSEA of DEPs ( C ; NES = −1.409, FDR = 0.143). The heatmap showed the top 20 DEPs ( D ), and the volcano diagram presented proteins with upregulation and downregulation in the CELSR2-KD group ( E ). F TOP/FOP flash luciferase assay showed a significant downregulation of Wnt/β-catenin signaling in the CELSR2-KD group. G , H Western blots of U87 MG cells showed the expression of CELSR2, β-catenin, GSK-3β, cyclin D1, p-β-catenin and p-GSK-3β in the CELSR2-KD group compared to the control group ( G ), with a significant upregulation of total GSK-3β, p-β-catenin and a significant downregulation of p-GSK-3β, total β-catenin and cyclin D1 ( H ). Data are represented as mean ± SEM. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001, triplicate experiments in each group, Student’s t -test.

Article Snippet: The sections were incubated with antibody against CELSR2 (1:200, NOVUS, NLS1943) overnight at 4 °C, followed by incubation with the secondary antibody for 1 h.

Techniques: Cell Culture, Transfection, Control, shRNA, Luciferase, Western Blot, Expressing

Journal: Cell Death & Disease

Article Title: Atypical cadherin CELSR2 acts as a therapeutic target for glioma through WNT3A/β-catenin signaling

doi: 10.1038/s41419-025-08116-8

Figure Lengend Snippet: A , B Cultured U87 MG cells transfected with CELSR2 -shRNA were treated without or with TWS119 ( CELSR2 KD or KD + TWS119), and cells without CELSR2 -shRNA transfection were used as the control. EDU labeling showed a decrease in EDU + cell density in the CELSR2-KD group compared to the control and the CELSR2 KD + TWS119 group. C , D Flow cytometry of cultured cells revealed an increase in the G0/G1 phase and a decrease in the S phase in the CELSR2-KD group compared to the control and the CELSR2 KD + TWS119 group. E , F Western blots showed lower expression of β-catenin and p-GSK-3β and higher expression of p-β-catenin and GSK-3β in the CELSR2-KD group compared to the control and the CELSR2 KD + TWS119 group. Scale bar is 50 μm in ( A ). Data are represented as mean ± SEM. * P < 0.05; ** P < 0.01; three independent experiments in each group; one-way ANOVA analysis of variance with Tukey’s multiple comparison.

Article Snippet: The sections were incubated with antibody against CELSR2 (1:200, NOVUS, NLS1943) overnight at 4 °C, followed by incubation with the secondary antibody for 1 h.

Techniques: Cell Culture, Transfection, shRNA, Control, Labeling, Flow Cytometry, Western Blot, Expressing, Comparison

Journal: Cell Death & Disease

Article Title: Atypical cadherin CELSR2 acts as a therapeutic target for glioma through WNT3A/β-catenin signaling

doi: 10.1038/s41419-025-08116-8

Figure Lengend Snippet: A , B Administration of WNT3A, WNT5A and WNT1 significantly enhanced cell proliferation of cultured glioma cells, as revealed by EDU staining. C , D In CELSR2-KD U87 MG cells, WNT5A and WNT1, but not WNT3A, significantly enhanced cell proliferation as identified by EDU labeling. E , F Flow cytometry showed a decrease in the G0/G1 ratio and an increase in the S phase in the control+WNT3A group (U87 MG cells treated with WNT3A) compared to the control group (U87 MG cells), and no significant change in the KD group (CELSR2-KD U87 MG cells) compared to the KD + WNT3A group (CELSR2-KD U87 MG cells treated with WNT3A). G , H Western blots showed a significant increase of β-catenin and cyclin D1, and a significant decrease in GSK-3β in the Ctrl+WNT3A group compared to the control group, and there was no significant protein change in the KD group compared to the KD + WNT3A group. Scale bar is 50 μm in ( A , C ). Data are represented as mean ± SEM. * P < 0.05; ** P < 0.01; *** P < 0.001; n = 3, Student’s t -test.

Article Snippet: The sections were incubated with antibody against CELSR2 (1:200, NOVUS, NLS1943) overnight at 4 °C, followed by incubation with the secondary antibody for 1 h.

Techniques: Cell Culture, Staining, Labeling, Flow Cytometry, Control, Western Blot

Journal: Cell Death & Disease

Article Title: Atypical cadherin CELSR2 acts as a therapeutic target for glioma through WNT3A/β-catenin signaling

doi: 10.1038/s41419-025-08116-8

Figure Lengend Snippet: A Schematic diagram of U87 MG cell injection in nude mice. B – E Nude mice were inoculated with U87 MG cells transfected with or without CELSR2 -shRNA (CELSR2-KD or control group). After one month of inoculation, tumor nodules appeared smaller ( B ) and their average weight was significantly lower ( C ) in the CELSR2-KD group compared to the control group. Dynamical measurement showed a significant decrease in tumor volume in the CELSR2-KD group compared to the control group ( D ), while animal body weight was comparable between the two groups ( E ). F , G Immunohistochemical staining of tumor tissues confirmed the downregulation of CELSR2 in the CELSR2-KD group. H In tumor nodules, there were a significant upregulation of p-β-catenin and GSK-3β and a significant downregulation of CELSR2 , β-catenin, p-GSK-3β and cyclinD1 in the CELSR2-KD group compared to the control group identified by RT-qPCR. I , J EDU labeling revealed a significant decrease in proliferating tumor cells in the CELSR2-KD group compared to the control group. Scale bar is 50 μm in ( F , I ). Data are represented as mean ± SEM. Two-way repeated measures ANOVA with Bonferroni’s post hoc correction in ( D , E ) and Student’s t -test in others, * P < 0.05, ** P < 0.01, *** P < 0.001, n = 5 in ( B – G ), ( I ) and ( J ), n = 3 in ( H ). A created with BioRender.

Article Snippet: The sections were incubated with antibody against CELSR2 (1:200, NOVUS, NLS1943) overnight at 4 °C, followed by incubation with the secondary antibody for 1 h.

Techniques: Injection, Transfection, shRNA, Control, Immunohistochemical staining, Staining, Quantitative RT-PCR, Labeling

Journal: Cell Death & Disease

Article Title: Atypical cadherin CELSR2 acts as a therapeutic target for glioma through WNT3A/β-catenin signaling

doi: 10.1038/s41419-025-08116-8

Figure Lengend Snippet: A Schematic diagram of MNPs-loaded CELSR2 -siRNA. B PB stain revealed MNPs-loaded CELSR2 -siRNA were ingested by cultured U87 MG cells. ( C ) CELSR2 mRNA levels were significantly decreased in the siRNA group and siRNA MNPs group compared to the control group. D , E EDU labeling showed a significant decrease in proliferating cells in the siRNA group and siRNA MNPs group. F , G Flow cytometry showed altered cell-cycle distribution (S phase decrease and G0/G1 increase in the siRNA group and siRNA MNPs group compared to the control). H Schematic diagram of nude mice inoculated with U87 MG cells and administration. I , J The tumor volume and average weight were significantly smaller in the siRNA MNPs group than in the control and MNPs groups. K , L The results of dynamic measurements of tumor volume and mouse body weight in live animals. M , N EDU labelling revealed a significant decrease in proliferating cells in the siRNA MNPs group compared to the control and MNPs groups. Scale bar is 100 μm in ( D ) and 50 μm in ( M ). Data are represented as mean ± SEM. Two-way repeated measures ANOVA with Bonferroni’s post hoc correction in ( K , L ) and one-way ANOVA analysis of variance with Tukey’s multiple comparison in others. * P < 0.05, ** P < 0.01, n = 3 in ( B – G ) and n = 8 in ( H – N ). ( A ) and ( H ) created with BioRender.

Article Snippet: The sections were incubated with antibody against CELSR2 (1:200, NOVUS, NLS1943) overnight at 4 °C, followed by incubation with the secondary antibody for 1 h.

Techniques: Staining, Cell Culture, Control, Labeling, Flow Cytometry, Comparison

Journal: Frontiers in Cell and Developmental Biology

Article Title: Celsr1 and Celsr2 exhibit distinct adhesive interactions and contributions to planar cell polarity

doi: 10.3389/fcell.2022.1064907

Figure Lengend Snippet: Generation of Celsr1 and Celsr2 loss-of function mutant mice by CRISPR/Cas9. (A) Schematic representation of Celsr1 and Celsr2 protein domains. The two proteins are 55% identical in amino acid sequence and have the same overall domain organization. (B) CRISPR-Cas9 targeting of Celsr1 and Celsr2 genomic loci. Guide RNAs were targeted to the sequence encoding the signal peptide for each of Celsr1 and Celsr2 . The resulting targeted alleles are shown with the ATG and signal sequence in purple font and deleted sequences highlighted in yellow. (C) Celsr1 −/− and wild type ( WT ) littermate at P12. Note curly tail and whorled hair pattern on the head of Celsr1 −/− homozygote. (D) Left and right paws of Celsr1 −/− and WT littermate at P12. Celsr1 −/− homozygotes exhibit prominent hair whorl on each paw. (E) Celsr1 −/− and WT littermate embryos at E15.5. Celsr1 −/− homozygotes display curly tail. (F) Western blot of epidermal lysates from WT and Celsr1 −/− P0-P3 backskins with anti-Celsr1 antibody. (G) Western blot of epidermal lysates from three individual WT and two individual Celsr2 −/− P0-P3 pups with anti-Celsr2 antibody. (H) Confocal immunofluorescence image of whole mount epidermis from E15.5 WT and Celsr1 −/− mutant embryos labeled with Celsr1 antibodies. Scale bars: 10 µm. (I) Quantification of Celsr1 mean fluorescence intensity in WT and Celsr1 −/− mutant epidermis (n = 3 skin regions from 4 different WT embryos and n = 3 skin regions from 3 different Celsr1 −/− embryos).

Article Snippet: Standard protocols were performed for western blot- proteins were resolved on a 7.5% SDS gel, transferred to a nitrocellulose membrane (Bio-Rad), and detected using primary antibodies against Celsr1 , Celsr2 (goat, R&D Systems, 1:200), and E-cadherin (rabbit, Cell Signaling, 1:250 or rat, ThermoFisher, 1:1000).

Techniques: Mutagenesis, CRISPR, Sequencing, Western Blot, Immunofluorescence, Labeling, Fluorescence

Journal: Frontiers in Cell and Developmental Biology

Article Title: Celsr1 and Celsr2 exhibit distinct adhesive interactions and contributions to planar cell polarity

doi: 10.3389/fcell.2022.1064907

Figure Lengend Snippet: Celsr1, but not Celsr2, is necessary for correct asymmetric orientation of developing hair follicles. (A) Average intensity projection of WT embryonic back skin at E15.5, labelled for P-cadherin (green) and Sox9 (magenta). White box denotes zoomed in region shown below, left. Average intensity projection of a typical WT hair follicle imaged at higher mag (below, right). Scale bars: 1000, 200, and 25 µm, respectively. Anterior is to the left. (B–D) As for (A) , except Celsr1 −/− , Celsr2 −/− and Celsr1 −/− ;Celsr2 −/− respectively. (E) Bar chart showing cumulative percentage of polarized (grey bar) vs. non-polarized (white bar) hair follicles in n = 3 E15.5 back skins from 3 different embryos. n in figure represents total number of follicles analyzed. Error bars = SEM. (F–H) As for (E) , except Celsr1 −/− , Celsr2 −/− and Celsr1 −/− ;Celsr2 −/− respectively. (I) Rose plot of polarized follicles in (E) showing the angle of orientation, with anterior = 0° and posterior = 180°. Shaded areas in bars represent relative contribution of each replicate (n = 3 backskins from 3 different embryos), with n in figure representing total number of polarized hair follicles analyzed. (J–L) As for (I) , except Celsr1 −/− , Celsr2 −/− and Celsr1 −/− ;Celsr2 −/− respectively.

Article Snippet: Standard protocols were performed for western blot- proteins were resolved on a 7.5% SDS gel, transferred to a nitrocellulose membrane (Bio-Rad), and detected using primary antibodies against Celsr1 , Celsr2 (goat, R&D Systems, 1:200), and E-cadherin (rabbit, Cell Signaling, 1:250 or rat, ThermoFisher, 1:1000).

Techniques:

Journal: Frontiers in Cell and Developmental Biology

Article Title: Celsr1 and Celsr2 exhibit distinct adhesive interactions and contributions to planar cell polarity

doi: 10.3389/fcell.2022.1064907

Figure Lengend Snippet: Loss of core PCP protein asymmetry in the epidermis of Celsr1 −/− and Celsr1 −/− ; Celsr2 −/− double mutants. (A–H) Representative planar views of the basal layer of the interfollicular epidermis at E15.5 showing Celsr1, Fz6 and Vangl2 distribution as detected by immunofluorescence. Anterior is to the left. Scale bar: 20 µm. Magnified areas below are overlaid with colored lines representing the axis (line angle) and magnitude (line length) of polarity. Quantification of polarity distributions are displayed below on circular histograms. (A-B′) Celsr1 (A) , Fz6 (B) and Vangl2 (B′) in WT embryos, n = 11,951 basal cells, 3 embryos. (C-D′) Celsr1 (C) , Fz6 (D) , and Vangl2 (D′) in Celsr1 −/− embryos, n = 11,629 basal cells, 3 embryos. (E-F′) Celsr1 (E) , Fz6 (F) and Vangl2 (F′) in Celsr2 −/− embryos, n = 12,099 basal cells, 3 embryos. (G-H′) Celsr1 (G) , Fz6 (H) and Vangl2 (H′) in Celsr1 −/− ; Celsr2 −/− embryos, n = 9,064 basal cells, 3 embryos.

Article Snippet: Standard protocols were performed for western blot- proteins were resolved on a 7.5% SDS gel, transferred to a nitrocellulose membrane (Bio-Rad), and detected using primary antibodies against Celsr1 , Celsr2 (goat, R&D Systems, 1:200), and E-cadherin (rabbit, Cell Signaling, 1:250 or rat, ThermoFisher, 1:1000).

Techniques: Immunofluorescence

Journal: Frontiers in Cell and Developmental Biology

Article Title: Celsr1 and Celsr2 exhibit distinct adhesive interactions and contributions to planar cell polarity

doi: 10.3389/fcell.2022.1064907

Figure Lengend Snippet: Celsr2 enriches at cell-cell junctions by homotypic interactions less efficiently than Celsr1. (A) Representative images of cell pairs expressing Celsr1-GFP, Celsr2-GFP or GFP-CAAX as indicated. Bottom panels show zoomed in junctional regions. Note the stronger enrichment of Celsr1-GFP at junctions compared to Celsr2-GFP and both isoforms are significantly more enriched at junctions compared to a non-junctional plasma membrane marker GFP-CAAX. Scale bars 20 µm (top panel) and 10 µm (bottom panel). (B) Plot of the junctional enrichment score (ratio of junctional mean intensity to the mean intensity of the cell pair). n = 32 Celsr1-GFP junctions, n = 43 Celsr2-GFP junctions, n = 60 GFP-CAAX junctions. Kolmogorov-Smirnov test p < 0.0001. Data pooled from two independent experiments where each experiment reflects the represented trend. (C) Fluorescence Recovery After Photobleaching (FRAP) of junctional Celsr1-GFP and Celsr2-GFP. Shown are representative images of the junctional region between cell pairs expressing Celsr1-GFP or Celsr2-GFP before and after bleaching as indicated. Bleached ROIs are marked by yellow arrowheads. (D) FRAP recovery plots. Shown is the normalized mean intensity with standard deviations of the bleach and recovery profiles plotted versus time for Celsr1-GFP (blue) and Celsr2-GFP (magenta) at junctions (in bold) and free cell edges that are not juxtaposed to a transfected cell (in lighter shade). (n = 36 ROIs for Celsr1 edge, 38 ROIs for Celsr2 edge, 78 ROIs for Celsr1 junctions and 75 ROIs for Celsr2 junctions). Data pooled from two independent experiments for cell edge measurements and three independent experiments for junction measurements. (E) Cell mixing experiment between cells expressing Celsr1-3xFLAG and Celsr2-GFP (top panels) or Celsr1-3xFLAG and Celsr1-GFP (bottom panels). Images show cell pairs forming heterotypic junctions. Celsr1-3xFLAG appears to enrich with both Celsr1-GFP and Celsr2-GFP, in trans , across cell-cell junctions. (F) Histogram depicting the frequency of Celsr1-3xFLAG: Celsr1-GFP and Celsr1-3xFLAG::Celsr2-GFP junctions across the range of junction enrichment ratios obtained for Celsr1-GFP and Celsr2-GFP, respectively. Inset shows box plot for the junction enrichment values of Celsr1-GFP and Celsr2-GFP. n = 56 Celsr1-GFP junctions and n = 64 Celsr2-GFP junctions. Kolmogorov-Smirnov test, p = 0.0004. Data pooled from two independent experiments.

Article Snippet: Standard protocols were performed for western blot- proteins were resolved on a 7.5% SDS gel, transferred to a nitrocellulose membrane (Bio-Rad), and detected using primary antibodies against Celsr1 , Celsr2 (goat, R&D Systems, 1:200), and E-cadherin (rabbit, Cell Signaling, 1:250 or rat, ThermoFisher, 1:1000).

Techniques: Expressing, Clinical Proteomics, Membrane, Marker, Fluorescence, Transfection

Journal: Frontiers in Cell and Developmental Biology

Article Title: Celsr1 and Celsr2 exhibit distinct adhesive interactions and contributions to planar cell polarity

doi: 10.3389/fcell.2022.1064907

Figure Lengend Snippet: Celsr2 recruits Fz6 and Vangl2 to keratinocyte junctions, similar to Celsr1. (A–C) Representative cell pair co-expressing Celsr1-GFP and Fz6-tdTomato (A) , tdTomato-Vangl2 (B) or a non-junctional membrane marker tdTomato-CAAX (C) . Arrowheads mark the junction between 2 cells and a magnified view of the junction is represented below the respective images. Scale bars = 20um. (D–F) Representative cell pair co-expressing Celsr2-GFP and Fz6-tdTomato (D) , tdTomato-Vangl2 (E) and tdTomato-CAAX (F) . Arrowheads mark the junction between 2 cells and a magnified view of the junction is represented below the respective images. Scale bars = 20 um. (G) Box plots depicting junction enrichment ratios for Fz6-tdTomato compared to tdTomato-CAAX when co-expressed with Celsr1-GFP or Celsr2-GFP (n = 33 for Celsr1-tdTomato CAAX, n = 49 for Celsr2-tdTomato-CAAX, n = 36 for Celsr1-Fz6-tdTomato, n = 66 for Celsr1-Fz6-tdTomato). (H) Box plots depicting junction enrichment values of Celsr1-GFP versus Celsr2-GFP in cells co-expressing Fz6-tdTomato. (I) Box plots depicting junction enrichment ratios for tdTomato-Vangl2 compared to tdTomato-CAAX when co-expressed with Celsr1-GFP or Celsr2-GFP (n = 66- Celsr1-tdTomatoCAAX, n = 87-Celsr2 tdTomatoCAAX, n = 63-Celsr1 tdTomatoCAAX, n = 67-Celsr2-tdTomatoCAAX). (J) Box plots depicting junction enrichment of Celsr1-GFP and Celsr2-GFP in cells expressing tdTomato-Vangl2. Data pooled from two independent experiments for Fz6-tdTomato and three independent experiments for tdTomato-Vangl2. Kolmogorov-Smirnov tests, **** p < 0.0001, ** p = 0.009.

Article Snippet: Standard protocols were performed for western blot- proteins were resolved on a 7.5% SDS gel, transferred to a nitrocellulose membrane (Bio-Rad), and detected using primary antibodies against Celsr1 , Celsr2 (goat, R&D Systems, 1:200), and E-cadherin (rabbit, Cell Signaling, 1:250 or rat, ThermoFisher, 1:1000).

Techniques: Expressing, Membrane, Marker

Journal: Frontiers in Cell and Developmental Biology

Article Title: Celsr1 and Celsr2 exhibit distinct adhesive interactions and contributions to planar cell polarity

doi: 10.3389/fcell.2022.1064907

Figure Lengend Snippet: Product information of key antibodies and reagents used in this study.

Article Snippet: Standard protocols were performed for western blot- proteins were resolved on a 7.5% SDS gel, transferred to a nitrocellulose membrane (Bio-Rad), and detected using primary antibodies against Celsr1 , Celsr2 (goat, R&D Systems, 1:200), and E-cadherin (rabbit, Cell Signaling, 1:250 or rat, ThermoFisher, 1:1000).

Techniques: Knock-Out

Journal: Frontiers in Cell and Developmental Biology

Article Title: Celsr1 and Celsr2 exhibit distinct adhesive interactions and contributions to planar cell polarity

doi: 10.3389/fcell.2022.1064907

Figure Lengend Snippet: Genotyping details for Celsr1 and Celsr2 including primer sequences and product sizes for WT and knockout animals.

Article Snippet: Standard protocols were performed for western blot- proteins were resolved on a 7.5% SDS gel, transferred to a nitrocellulose membrane (Bio-Rad), and detected using primary antibodies against Celsr1 , Celsr2 (goat, R&D Systems, 1:200), and E-cadherin (rabbit, Cell Signaling, 1:250 or rat, ThermoFisher, 1:1000).

Techniques: Knock-Out, Western Blot, Plasmid Preparation, Software

Journal: BMC Cancer

Article Title: Identification of CELSR2 as a novel prognostic biomarker for hepatocellular carcinoma

doi: 10.1186/s12885-020-06813-5

Figure Lengend Snippet: Gene and protein expression profiles of CELSR2 in human normal and tumor samples. a CELSR2 mRNA expression data from the GTEx project in the Human Protein Atlas. b CELSR2 gene expression in common human tumor tissues in the Human Protein Atlas. c CELSR2 protein expression overview in human tumor tissues in the Human Protein Atlas. d CELSR2 mRNA expression overview in human cancer cell lines in the Human Protein Atlas

Article Snippet: HepG2 and Hep3B hepatoma cell lines in 6-well plates were transfected with CELSR2 small inference RNA (siRNA) and NC by Lipofectamine 2000 (Invitrogen), which were designed and synthesized by RiboBio company (Guangzhou, China) at a final concentration of 15 nM.

Techniques: Expressing, Gene Expression

Journal: BMC Cancer

Article Title: Identification of CELSR2 as a novel prognostic biomarker for hepatocellular carcinoma

doi: 10.1186/s12885-020-06813-5

Figure Lengend Snippet: Gene expression profiles of CELSR2 in the HCCDB database. a Radar map of CELSR2 overall expression among different types of tissues. b The differential expression of CELSR2 in different liver cancer datasets (HCCDB1, HCCDB3, HCCDB4, HCCDB6, HCCDB7, HCCDB13, HCCDB15, HCCDB17 and HCCDB18) suggests that CELSR2 expression is much higher in HCC tissues than in adjacent liver tissues. The coexpression networks of CELSR2 in HCC tissues ( c ), adjacent liver tissues ( d ) and normal tissues from the GTEx project ( e ) showed that different liver tissues expressed different coexpression networks

Article Snippet: HepG2 and Hep3B hepatoma cell lines in 6-well plates were transfected with CELSR2 small inference RNA (siRNA) and NC by Lipofectamine 2000 (Invitrogen), which were designed and synthesized by RiboBio company (Guangzhou, China) at a final concentration of 15 nM.

Techniques: Gene Expression, Expressing, Quantitative Proteomics

Journal: BMC Cancer

Article Title: Identification of CELSR2 as a novel prognostic biomarker for hepatocellular carcinoma

doi: 10.1186/s12885-020-06813-5

Figure Lengend Snippet: The mRNA expression of CELSR2 in the UALCAN database. a The mRNA expression level of CELSR2 was significantly higher in cancer tissues than in normal tissues. b The expression level of CELSR2 in female patients was higher than that in healthy people or in male patients. c Patients (age > 21 years old) commonly had higher gene expression than young healthy people. The expression level of CELSR2 was positively correlated with patient weight ( d ), tumor stage ( e ) and tumor grade ( f ) in HCC patients. *** represents p < 0.001

Article Snippet: HepG2 and Hep3B hepatoma cell lines in 6-well plates were transfected with CELSR2 small inference RNA (siRNA) and NC by Lipofectamine 2000 (Invitrogen), which were designed and synthesized by RiboBio company (Guangzhou, China) at a final concentration of 15 nM.

Techniques: Expressing, Gene Expression

Journal: BMC Cancer

Article Title: Identification of CELSR2 as a novel prognostic biomarker for hepatocellular carcinoma

doi: 10.1186/s12885-020-06813-5

Figure Lengend Snippet: Gene expression of CELSR2 and its prognostic value in HCC patients. a Prognostic value of the CELSR2 level in HCC patients from the Human Protein Atlas. b Representative immunohistochemistry (IHC) images from the Human Protein Atlas with the CELSR2 antibody: HPA013952, cancerous tissue had higher staining than that in normal liver tissue. c High CELSR2 mRNA expression was significantly related to poor OS in HCC patients from the Kaplan-Meier plotter

Article Snippet: HepG2 and Hep3B hepatoma cell lines in 6-well plates were transfected with CELSR2 small inference RNA (siRNA) and NC by Lipofectamine 2000 (Invitrogen), which were designed and synthesized by RiboBio company (Guangzhou, China) at a final concentration of 15 nM.

Techniques: Gene Expression, Immunohistochemistry, Staining, Expressing

Journal: BMC Cancer

Article Title: Identification of CELSR2 as a novel prognostic biomarker for hepatocellular carcinoma

doi: 10.1186/s12885-020-06813-5

Figure Lengend Snippet: Genetic mutations of CELSR2 and its associations with OS in HCC patients (cBioPortal). a Oncoprint of CELSR2 alterations in HCC. The overview of genomic alterations showed that the alteration rate of CELSR2 was 8%. b Mutation types of CELSR2 gene in HCC patients. c Network view of CELSR2 and its altered neighboring genes in HCC. d GO functional enrichment and KEGG pathway analyses of CELSR2 and its frequently altered neighboring genes

Article Snippet: HepG2 and Hep3B hepatoma cell lines in 6-well plates were transfected with CELSR2 small inference RNA (siRNA) and NC by Lipofectamine 2000 (Invitrogen), which were designed and synthesized by RiboBio company (Guangzhou, China) at a final concentration of 15 nM.

Techniques: Mutagenesis, Functional Assay

Journal: BMC Cancer

Article Title: Identification of CELSR2 as a novel prognostic biomarker for hepatocellular carcinoma

doi: 10.1186/s12885-020-06813-5

Figure Lengend Snippet: Genes differentially expressed in correlation with CELSR2 in HCC (LinkedOmics). a Volcano plot showing genes correlated with CELSR2 through Pearson’s test analysis. b, c Heat maps showing the top 50 genes positively and negatively correlated with CELSR2in HCC; red (positive), green (negative). The significantly enriched GO annotations ( d ) cellular components, ( e ) biological processes and ( f ) molecular functions), KEGG pathways ( g ), kinase ( h ), miRNA ( i ) and transcription factor ( j ) targets of CELSR2 coexpressed genes in HCC were analyzed using GSEA

Article Snippet: HepG2 and Hep3B hepatoma cell lines in 6-well plates were transfected with CELSR2 small inference RNA (siRNA) and NC by Lipofectamine 2000 (Invitrogen), which were designed and synthesized by RiboBio company (Guangzhou, China) at a final concentration of 15 nM.

Techniques:

Journal: BMC Cancer

Article Title: Identification of CELSR2 as a novel prognostic biomarker for hepatocellular carcinoma

doi: 10.1186/s12885-020-06813-5

Figure Lengend Snippet: Expression and biological function of CELSR2 in HCC. a The mRNA and protein levels of CELSR2 from cultured cells by qPCR and western blot assays. The expression of CELSR2 protein was normalized to GAPDH. b, c CELSR2 knockdown prohibited the proliferation rate of hepatoma cells. d-k CELSR2 knockdown reduced cell invasion in both HepG2 and Hep3B cells. l Representative cases of high and low CELSR2 expression by immunohistochemistry staining. m Semi-quantitative analysis of CELSR2 protein expression between cancerous specimens and non-cancerous parts. * represents p < 0.05, *** represents p < 0.01, *** represents p < 0.001

Article Snippet: HepG2 and Hep3B hepatoma cell lines in 6-well plates were transfected with CELSR2 small inference RNA (siRNA) and NC by Lipofectamine 2000 (Invitrogen), which were designed and synthesized by RiboBio company (Guangzhou, China) at a final concentration of 15 nM.

Techniques: Expressing, Cell Culture, Western Blot, Knockdown, Immunohistochemistry, Staining

Journal: BMC Cancer

Article Title: Identification of CELSR2 as a novel prognostic biomarker for hepatocellular carcinoma

doi: 10.1186/s12885-020-06813-5

Figure Lengend Snippet: Prognostic factors associated with recurrence-free survival

Article Snippet: HepG2 and Hep3B hepatoma cell lines in 6-well plates were transfected with CELSR2 small inference RNA (siRNA) and NC by Lipofectamine 2000 (Invitrogen), which were designed and synthesized by RiboBio company (Guangzhou, China) at a final concentration of 15 nM.

Techniques:

Journal: BMC Cancer

Article Title: Identification of CELSR2 as a novel prognostic biomarker for hepatocellular carcinoma

doi: 10.1186/s12885-020-06813-5

Figure Lengend Snippet: Prognostic factors associated with overall survival

Article Snippet: HepG2 and Hep3B hepatoma cell lines in 6-well plates were transfected with CELSR2 small inference RNA (siRNA) and NC by Lipofectamine 2000 (Invitrogen), which were designed and synthesized by RiboBio company (Guangzhou, China) at a final concentration of 15 nM.

Techniques: