Journal: Cells

Article Title: CHRDL1 Regulates Stemness in Glioma Stem-like Cells

doi: 10.3390/cells11233917

Figure Lengend Snippet: CHRDL1 depletion blocks stemness in GSCs. ( A ) Western Blot of NCH644 and GS-5 GSCs after stable transduction with a non-targeting shRNA (shCtrl) or specific shRNA targeting CHRDL1 (shCHRDL1) and ( B ) Quantification of several independent experiments. ( C , E , F ) Log-fraction plots of the limited dilution model of data from ( C ) NCH644 shCtrl (black) and NCH644 shCHRDL1 (red) GSCs 7 days after seeding the cell in a dilution series from 1024 to 8 cells and analyzing the data using ELDA software . ( D ) Representative microphotographs of NCH644 and GS-5 with and without CRHDL1-depletion. Scale bar: 200 µm. ( E , F ) Data from GS-5 GSCs ( E ) 7 days and ( F ) 14 days after seeding of 2048 to 16 cells. **: p < 0.01, ***: p < 0.001 ****: p < 0.0001.

Article Snippet: The following primary antibodies and dilutions were used: CHRDL1 (#AF1808, R&D Systems, Wiesbaden-Nordenstadt, Germany) 1:2000; GAPDH (#CB1001, Calbiochem, Darmstadt, Germany) 1:20,000; OLIG2 (#AF2418, R&D Biosystems) 1:5000; SOX2 (#MAB2018, R&D Systems) 1:5000; SOX9 (#ab185966, Abcam, Cambridge, UK); SMAD1 [(#6944, Cell Signaling Technology (CST), Danvers, MA, USA)] 1:1000; SMAD5 [(#12534, CST)] 1:1000; pSMAD1/5 [(#9516, CST)] 1:1000.

Techniques: Western Blot, Transduction, shRNA, Software

Journal: Cells

Article Title: CHRDL1 Regulates Stemness in Glioma Stem-like Cells

doi: 10.3390/cells11233917

Figure Lengend Snippet: Cell cycle distribution of GSCs after CHRDL1-depletion. ( A , B ) Stacked bar graph of ( A ) NCH644 and ( B ) GS-5 after stable transduction with a non-targeting shRNA (shCtrl) or specific shRNA targeting CHRDL1 (shCHRDL1). After 24 h of seeding the cells, the cell cycle distribution was analyzed. For NCH644 GSCs a significant increase in cells in ( A ′) G1-phase with a concomitant significant decrease in ( A ″) S- and ( A ‴) G2/M-phases was observed. The experiment was performed using biological quadruplicates. * p < 0.05; ** p < 0.01; ns: not significant; unpaired t test.

Article Snippet: The following primary antibodies and dilutions were used: CHRDL1 (#AF1808, R&D Systems, Wiesbaden-Nordenstadt, Germany) 1:2000; GAPDH (#CB1001, Calbiochem, Darmstadt, Germany) 1:20,000; OLIG2 (#AF2418, R&D Biosystems) 1:5000; SOX2 (#MAB2018, R&D Systems) 1:5000; SOX9 (#ab185966, Abcam, Cambridge, UK); SMAD1 [(#6944, Cell Signaling Technology (CST), Danvers, MA, USA)] 1:1000; SMAD5 [(#12534, CST)] 1:1000; pSMAD1/5 [(#9516, CST)] 1:1000.

Techniques: Transduction, shRNA

Journal: Cells

Article Title: CHRDL1 Regulates Stemness in Glioma Stem-like Cells

doi: 10.3390/cells11233917

Figure Lengend Snippet: Point-plots of Taqman-based gene expression of NCH644 GSCs after stable transduction with a non-targeting shRNA (shCtrl) or specific shRNA targeting CHRDL1 (shCHRDL1) and measurement of ( A ) CHRDL1 , ( B ) OLIG2 , ( C ) SOX2 , ( D ) SOX9 , ( E ) ZEB1 , ( F ) ZEB2 , ( G ) CD44 , ( H ) NES , ( I ) GFAP , ( J ) NEFL , ( K ) MAP2 and ( L ) RBFOX3 expression. The data are the summary of three experiments performed in triplicates. * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001; ns: not significant; unpaired t test with Welch’s correction; white dot: shCtrl, red dot: shCHRDL1.

Article Snippet: The following primary antibodies and dilutions were used: CHRDL1 (#AF1808, R&D Systems, Wiesbaden-Nordenstadt, Germany) 1:2000; GAPDH (#CB1001, Calbiochem, Darmstadt, Germany) 1:20,000; OLIG2 (#AF2418, R&D Biosystems) 1:5000; SOX2 (#MAB2018, R&D Systems) 1:5000; SOX9 (#ab185966, Abcam, Cambridge, UK); SMAD1 [(#6944, Cell Signaling Technology (CST), Danvers, MA, USA)] 1:1000; SMAD5 [(#12534, CST)] 1:1000; pSMAD1/5 [(#9516, CST)] 1:1000.

Techniques: Gene Expression, Transduction, shRNA, Expressing

Journal: Cells

Article Title: CHRDL1 Regulates Stemness in Glioma Stem-like Cells

doi: 10.3390/cells11233917

Figure Lengend Snippet: Point-plots of Taqman-based gene expression of GS-5 GSCs after stable transduction with a non-targeting shRNA (shCtrl) or specific shRNA targeting CHRDL1 (shCHRDL1) and measurement of ( A ) CHRDL1 , ( B ) OLIG2 , ( C ) SOX2 , ( D ) SOX9 , ( E ) ZEB1 , ( F ) ZEB2 , ( G ) CD44 , ( H ) NES , ( I ) GFAP , ( J ) NEFL , ( K ) MAP2 and ( L ) RBFOX3 expression. The data are the summary of three experiments performed in triplicates. * p < 0.05; *** p < 0.001; **** p < 0.0001; ns: not significant; unpaired t test with Welch’s correction; white dot: shCtrl, red dot: shCHRDL1.

Article Snippet: The following primary antibodies and dilutions were used: CHRDL1 (#AF1808, R&D Systems, Wiesbaden-Nordenstadt, Germany) 1:2000; GAPDH (#CB1001, Calbiochem, Darmstadt, Germany) 1:20,000; OLIG2 (#AF2418, R&D Biosystems) 1:5000; SOX2 (#MAB2018, R&D Systems) 1:5000; SOX9 (#ab185966, Abcam, Cambridge, UK); SMAD1 [(#6944, Cell Signaling Technology (CST), Danvers, MA, USA)] 1:1000; SMAD5 [(#12534, CST)] 1:1000; pSMAD1/5 [(#9516, CST)] 1:1000.

Techniques: Gene Expression, Transduction, shRNA, Expressing

Journal: Cells

Article Title: CHRDL1 Regulates Stemness in Glioma Stem-like Cells

doi: 10.3390/cells11233917

Figure Lengend Snippet: CHRDL1-depletion reduces stemness-associated marker protein expression. Western Blots of GS-5 GSCs after stable transduction with a non-targeting shRNA (shCtrl) or specific shRNA targeting CHRDL1 (shCHRDL1) and measurement of the protein expression of OLIG2, SOX9, and SOX2. CHRDL1 was detected to validate efficient depletion and GAPDH served as a housekeeping protein.

Article Snippet: The following primary antibodies and dilutions were used: CHRDL1 (#AF1808, R&D Systems, Wiesbaden-Nordenstadt, Germany) 1:2000; GAPDH (#CB1001, Calbiochem, Darmstadt, Germany) 1:20,000; OLIG2 (#AF2418, R&D Biosystems) 1:5000; SOX2 (#MAB2018, R&D Systems) 1:5000; SOX9 (#ab185966, Abcam, Cambridge, UK); SMAD1 [(#6944, Cell Signaling Technology (CST), Danvers, MA, USA)] 1:1000; SMAD5 [(#12534, CST)] 1:1000; pSMAD1/5 [(#9516, CST)] 1:1000.

Techniques: Marker, Expressing, Western Blot, Transduction, shRNA

Journal: Cells

Article Title: CHRDL1 Regulates Stemness in Glioma Stem-like Cells

doi: 10.3390/cells11233917

Figure Lengend Snippet: CHRDL1-depletion activates BMP4-signaling. ( A , B ) Western Blot of ( A ) NCH644 treated with 20 ng/mL recombinant human BMP4 for 24 or 48 h or ( B ) NCH644 shCtrl and shCHRDL1. ( C , D ) Sphere formation assay of ( C ) NCH644 and ( D ) GS-5 shCtrl or shCHRDL1 GSCs and treatment with 25 ng/mL recombinant BMP4 or solvent for 7 days after seeding of 500 and 1000 cells per well. * p < 0.05; **** p < 0.0001; two-way-ANOVA with Sidak’s multiple comparison tests; white dot: shCtrl, red dot: shCHRDL1.

Article Snippet: The following primary antibodies and dilutions were used: CHRDL1 (#AF1808, R&D Systems, Wiesbaden-Nordenstadt, Germany) 1:2000; GAPDH (#CB1001, Calbiochem, Darmstadt, Germany) 1:20,000; OLIG2 (#AF2418, R&D Biosystems) 1:5000; SOX2 (#MAB2018, R&D Systems) 1:5000; SOX9 (#ab185966, Abcam, Cambridge, UK); SMAD1 [(#6944, Cell Signaling Technology (CST), Danvers, MA, USA)] 1:1000; SMAD5 [(#12534, CST)] 1:1000; pSMAD1/5 [(#9516, CST)] 1:1000.

Techniques: Western Blot, Recombinant, Tube Formation Assay, Solvent, Comparison

Journal: Cells

Article Title: CHRDL1 Regulates Stemness in Glioma Stem-like Cells

doi: 10.3390/cells11233917

Figure Lengend Snippet: CHRDL1-depletion radiosensitizes GSCs. ( A ) 53BP1 foci assay 24 h after irradiation of NCH644 shCtrl or NCH644 shCHRDL1 GSCs with the indicated doses. ( B ) Sphere formation of NCH644 shCtrl or shCHRDL1 cells 7 days after seeding of single cells and irradiation as indicated. ( C ) Representative microphotographs of NCH644 shCtrl or NCH644 shCRHDL1 without irradiation (0 Gy) or after irradiation with 6 Gy; scale bar: 200 µm. * p < 0.05; ** p < 0.01; **** p < 0.0001; two-way-ANOVA with Sidak’s multiple comparison tests. ( D ) Sphere formation of NCH644 treated with solvent or recombinant human BMP4 (25 ng/mL) immediately prior to irradiation with a dose of 4 or 6 Gy. Sphere area was determined 7 days after treatment. * p < 0.05; **** p < 0.0001; Kruskal—Wallis test with Dunn’s multiple comparisons test.

Article Snippet: The following primary antibodies and dilutions were used: CHRDL1 (#AF1808, R&D Systems, Wiesbaden-Nordenstadt, Germany) 1:2000; GAPDH (#CB1001, Calbiochem, Darmstadt, Germany) 1:20,000; OLIG2 (#AF2418, R&D Biosystems) 1:5000; SOX2 (#MAB2018, R&D Systems) 1:5000; SOX9 (#ab185966, Abcam, Cambridge, UK); SMAD1 [(#6944, Cell Signaling Technology (CST), Danvers, MA, USA)] 1:1000; SMAD5 [(#12534, CST)] 1:1000; pSMAD1/5 [(#9516, CST)] 1:1000.

Techniques: Irradiation, Comparison, Solvent, Recombinant

Journal: Cells

Article Title: CHRDL1 Regulates Stemness in Glioma Stem-like Cells

doi: 10.3390/cells11233917

Figure Lengend Snippet: CHRDL1 is associated with worse survival in glioma and ( A ) An unfavourable prognosis in ( B ) urothelial and ( C ) renal cancer. According to data from the human protein atlas (proteinatlas.org). * p < 0.05; *** p < 0.001; log-rank p -value for Kaplan–Meier plot. Image Credit: Human Protein Atlas.

Article Snippet: The following primary antibodies and dilutions were used: CHRDL1 (#AF1808, R&D Systems, Wiesbaden-Nordenstadt, Germany) 1:2000; GAPDH (#CB1001, Calbiochem, Darmstadt, Germany) 1:20,000; OLIG2 (#AF2418, R&D Biosystems) 1:5000; SOX2 (#MAB2018, R&D Systems) 1:5000; SOX9 (#ab185966, Abcam, Cambridge, UK); SMAD1 [(#6944, Cell Signaling Technology (CST), Danvers, MA, USA)] 1:1000; SMAD5 [(#12534, CST)] 1:1000; pSMAD1/5 [(#9516, CST)] 1:1000.

Techniques:

Journal: Diabetes

Article Title: BMP4 and BMP Antagonists Regulate Human White and Beige Adipogenesis.

doi: 10.2337/db14-1127

Figure Lengend Snippet: Figure 3 - Silencing the BMP4 inhibitor CHRDL1 in preadipocytes elicits negative effect on adipogenesis.

Article Snippet: Over expression of CHRDL1 7 To overexpress CHRDL1, cells were transfected with a myc-DDK-tagged ORF clone of CHRDL1 obtained from Origene (TrueORF Gold RC202635, BioNordika Sweden) using Lipofectamine 2000 (Life Technologies, Stockholm, Sweden) according to the manufacturers protocol.

Techniques:

Journal: Frontiers in Oncology

Article Title: Chorordin-like 1 inhibits pancreatic cancer cell migration and invasion: involvement of the BMP4/SMAD pathway

doi: 10.3389/fonc.2025.1633464

Figure Lengend Snippet: Decreased expression of CHRDL1 in pancreatic cancer. (A) Data-based prediction of CHRDL1 expression in pancreatic cancer cells, two-tailed Student’s t-test. (B) Association between CHRDL1 and disease-free survival (DFS). (C-E) The efficiency of CHRDL1 overexpression in the pancreatic cancer cell lines PANC-1 and SW1990 was verified using quantitative real-time PCR (RT-qPCR) and Western blot analysis (n = 6, * P < 0.05 vs. hTERT-HPNE, one-way ANOVA followed by Sidak’s multiple comparisons test for multi-group). (F, G) IHC analysis of CHRDL1 in tumor and adjacent tissues (n = 6, * P < 0.05 vs. adjacent, two-tailed Student’s t-test).

Article Snippet: The EGFP and CHRDL1 genes (OriGene, #RC202635) were subcloned into the lentiviral vector pLV-EF1a-IRES-Ametrine.

Techniques: Expressing, Two Tailed Test, Over Expression, Real-time Polymerase Chain Reaction, Quantitative RT-PCR, Western Blot

Journal: Frontiers in Oncology

Article Title: Chorordin-like 1 inhibits pancreatic cancer cell migration and invasion: involvement of the BMP4/SMAD pathway

doi: 10.3389/fonc.2025.1633464

Figure Lengend Snippet: Overexpression of CHRDL1 inhibits malignant biological behaviors of PANC-1 and SW1990 cells. (A, B) Representative images of wound healing at 0 and 24 h in CHRDL1 transduced PANC-1 cells, along with statistical results of migration distance percentages for each group (Scale bars: 100 μm). (C, D) Migration and invasion assays of transfected PANC-1 cells, showing representative images and corresponding statistical analysis for each group (scale bars, upper: 50 μm, lower: 20 μm). (E, F) Representative images of wound healing at 0 and 24 h in CHRDL1 transduced SW1990 cells, along with statistical results of migration distance percentages for each group (Scale bars: 100 μm). (G, H) Migration and invasion assays of transfected SW1990 cells, showing representative images and corresponding statistical analysis for each group (scale bars, upper: 50 μm, lower: 20 μm). n = 6 in each group, * P < 0.05 vs. control, one-way ANOVA followed by Sidak’s multiple comparisons test for multi-group.

Article Snippet: The EGFP and CHRDL1 genes (OriGene, #RC202635) were subcloned into the lentiviral vector pLV-EF1a-IRES-Ametrine.

Techniques: Over Expression, Migration, Transfection, Control

Journal: Frontiers in Oncology

Article Title: Chorordin-like 1 inhibits pancreatic cancer cell migration and invasion: involvement of the BMP4/SMAD pathway

doi: 10.3389/fonc.2025.1633464

Figure Lengend Snippet: Overexpression of CHRDL1 inhibits BMP4-Induced SMAD phosphorylation in pancreatic cancer cells. (A, B) p-SMAD 1/5/9, t-SMAD 1/5/9, RUNX2 and CHRDL1 protein levels in PANC-1 cells were detected using Western blots. The immunoblots were calculated by densitometric analysis using GAPDH as the internal reference. (C, D) p-SMAD 1/5/9, t-SMAD 1/5/9, RUNX2 and CHRDL1 protein levels in SW1990 cells were detected using Western blots. The immunoblots were calculated by densitometric analysis using GAPDH as the internal reference. n = 6 in each group, * P < 0.05 vs. OE-NC, # P < 0.05 rhBMP4+OE-NC, one-way ANOVA followed by Sidak’s multiple comparisons test for multi-group.

Article Snippet: The EGFP and CHRDL1 genes (OriGene, #RC202635) were subcloned into the lentiviral vector pLV-EF1a-IRES-Ametrine.

Techniques: Over Expression, Phospho-proteomics, Western Blot

Journal: Frontiers in Oncology

Article Title: Chorordin-like 1 inhibits pancreatic cancer cell migration and invasion: involvement of the BMP4/SMAD pathway

doi: 10.3389/fonc.2025.1633464

Figure Lengend Snippet: CHRDL1 attenuates BMP4-induced malignant biological behaviors of pancreatic cancer cells. (A, B) Representative wound healing images of CHRDL1-transduced PANC-1 cells treated with rhBMP4 at 0 and 12 hours, along with statistical results of migration distance percentages for each group (Scale bars: 100 μm). (C, D) Migration and invasion assays of transfected PANC-1 cells treated with rhBMP4, showing representative images and corresponding statistical analysis for each group (scale bars, upper: 50 μm, lower: 20 μm). (E, F) Representative wound healing images of CHRDL1-transduced SW1990 cells treated with rhBMP4 at 0 and 12 hours, along with statistical results of migration distance percentages for each group (Scale bars: 100 μm). (G, H) Migration and invasion assays of transfected SW1990 cells treated with rhBMP4, showing representative images and corresponding statistical analysis for each group (scale bars, upper: 50 μm, lower: 20 μm).n = 6 in each group, *P < 0.05 vs. OE-NC, one-way ANOVA followed by Sidak’s multiple comparisons test for multi-group.

Article Snippet: The EGFP and CHRDL1 genes (OriGene, #RC202635) were subcloned into the lentiviral vector pLV-EF1a-IRES-Ametrine.

Techniques: Migration, Transfection

Journal: Frontiers in Oncology

Article Title: Chorordin-like 1 inhibits pancreatic cancer cell migration and invasion: involvement of the BMP4/SMAD pathway

doi: 10.3389/fonc.2025.1633464

Figure Lengend Snippet: CHRDL1 inhibits pancreatic cancer metastasis in vivo. (A, B) Representative lung images of BALB/c nude mice at 9 weeks after tail vein injection with OE-NC/OE-CHRDL1 stably transfected PANC-1 cells (left) or SW1990 cells (right). (C, D) Lung weights of BALB/c nude mice at 9 weeks after tail vein injection with OE-NC/OE-CHRDL1 stably transfected PANC-1 cells (left) or SW1990 cells (right). (E-H) H&E-stained lung sections and corresponding quantitative analysis of tumor area/total lung area from mice injected with OE-NC/OE-CHRDL1 stably transfected PANC-1 cells (left) or SW1990 cells (right). Scale bars, upper: 2 mm, lower: 100 μm. n = 6 in each group, * P < 0.05 vs. OE-NC, two-tailed Student’s t-test.

Article Snippet: The EGFP and CHRDL1 genes (OriGene, #RC202635) were subcloned into the lentiviral vector pLV-EF1a-IRES-Ametrine.

Techniques: In Vivo, Injection, Stable Transfection, Transfection, Staining, Two Tailed Test

Journal: Frontiers in Oncology

Article Title: Chorordin-like 1 inhibits pancreatic cancer cell migration and invasion: involvement of the BMP4/SMAD pathway

doi: 10.3389/fonc.2025.1633464

Figure Lengend Snippet: Schematic illustration of the mechanism whereby CHRDL1 regulates pancreatic cancer. In general, CHRDL1 functions as a BMP antagonist that desensitizes pancreatic cancer cells to BMP-4, thereby inhibiting BMP-mediated SMAD1/5/9 signaling activation. Ultimately, CHRDL1 suppresses malignant behaviors of pancreatic cancer cells and impedes tumor progression in vivo .

Article Snippet: The EGFP and CHRDL1 genes (OriGene, #RC202635) were subcloned into the lentiviral vector pLV-EF1a-IRES-Ametrine.

Techniques: Activation Assay, In Vivo

Journal: Frontiers in Oncology

Article Title: Chordin-Like 1 Regulates Epithelial-to-Mesenchymal Transition and Metastasis via the MAPK Signaling Pathway in Oral Squamous Cell Carcinoma

doi: 10.3389/fonc.2022.862751

Figure Lengend Snippet: The expression of CHRDL1 was downregulated in OSCC tissue and cell lines. (A) Relative mRNA expression of CHRDL1 in OSCC tissues ( n = 201) and normal tissues ( n = 18) from the TCGA database. (B) The relative mRNA expression of CHRDL1 in 30 specimens of OSCC and adjacent tissues was determined by qRT-PCR. (C) The relative mRNA expression of CHRDL1 in normal human oral keratinocytes (HOK) and the OSCC cell line CAL27 was determined by RT-qPCR. CHRDL1 mRNA expression was normalized to GAPDH. (D, E) IHC analysis of CHRDL1 in tumor and adjacent tissues ( n = 5). Error bars represent the standard deviation. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: Tissue sections were further incubated with primary antibody against CHRDL1 (1:150 dilution, Zen-Bio) at 4°C for 12 h and then washed and incubated with secondary antibody (1:100 dilution, ZSJB-Bio) at 37°C for 1 h. Finally, slides were incubated with diaminobenzidine (DAB, ZSJB-Bio) and counterstained with hematoxylin.

Techniques: Expressing, Quantitative RT-PCR, Standard Deviation

Journal: Frontiers in Oncology

Article Title: Chordin-Like 1 Regulates Epithelial-to-Mesenchymal Transition and Metastasis via the MAPK Signaling Pathway in Oral Squamous Cell Carcinoma

doi: 10.3389/fonc.2022.862751

Figure Lengend Snippet: Overexpression of CHRDL1 inhibits CAL27 cell migration, invasion, and adhesion. (A) Relative mRNA expression of CHRDL1 in CAL27 cells after lentivirus transfection. (B, C) CHRDL1 protein expression in CAL27 cells after lentivirus transfection was identified by Western blot and quantitatively analyzed. (D, E) Representative images of wound-healing assays of transfected CAL27 cells at 0 and 24 h and statistical results from each group. (F, G) Migration and invasion assays of transfected CAL27 cells. Representative images of cell migration and invasion in each group are shown in (F) . Statistical analyses of cell migration and invasion are shown in (G) . (H, I) Cell adhesion assay of transfected CAL27 cells. Representative fluorescence images from each group (H) and statistical analyses of the adherence rate (I) . Error bars represent the standard deviation. ** p < 0.01, *** p < 0.001.

Article Snippet: Tissue sections were further incubated with primary antibody against CHRDL1 (1:150 dilution, Zen-Bio) at 4°C for 12 h and then washed and incubated with secondary antibody (1:100 dilution, ZSJB-Bio) at 37°C for 1 h. Finally, slides were incubated with diaminobenzidine (DAB, ZSJB-Bio) and counterstained with hematoxylin.

Techniques: Over Expression, Migration, Expressing, Transfection, Western Blot, Cell Adhesion Assay, Fluorescence, Standard Deviation

Journal: Frontiers in Oncology

Article Title: Chordin-Like 1 Regulates Epithelial-to-Mesenchymal Transition and Metastasis via the MAPK Signaling Pathway in Oral Squamous Cell Carcinoma

doi: 10.3389/fonc.2022.862751

Figure Lengend Snippet: CHRDL1 knockdown promotes CAL27 cell migration, invasion, and adhesion. (A) Relative mRNA expression of CHRDL1 in CAL27 cells after lentivirus transfection. (B, C) CHRDL1 protein expression in CAL27 cells after lentivirus transfection was evaluated by Western blot and quantitatively analyzed. (D, E) Representative images of wound-healing assays of transfected CAL27 cells at 0 and 24 h and statistical analyses of each group. (F, G) Migration and invasion assays of transfected CAL27 cells. Representative images of cell migration and invasion in each group are shown in (F) . Statistical analyses of cell migration and invasion are shown in (G) . (H, I) Cell adhesion assay of transfected CAL27 cells. Representative fluorescence images of each group (H) and statistical analyses of the adherence rate (I) . Error bars represent the standard deviation. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: Tissue sections were further incubated with primary antibody against CHRDL1 (1:150 dilution, Zen-Bio) at 4°C for 12 h and then washed and incubated with secondary antibody (1:100 dilution, ZSJB-Bio) at 37°C for 1 h. Finally, slides were incubated with diaminobenzidine (DAB, ZSJB-Bio) and counterstained with hematoxylin.

Techniques: Knockdown, Migration, Expressing, Transfection, Western Blot, Cell Adhesion Assay, Fluorescence, Standard Deviation

Journal: Frontiers in Oncology

Article Title: Chordin-Like 1 Regulates Epithelial-to-Mesenchymal Transition and Metastasis via the MAPK Signaling Pathway in Oral Squamous Cell Carcinoma

doi: 10.3389/fonc.2022.862751

Figure Lengend Snippet: Depletion of CHRDL1 induces the EMT phenotype in CAL27 cells. (A) Representative fluorescence images of sh-NC/sh1-CHRDL1 CAL27 cells under low (A) and high (b) cell density conditions. (c) Rhodamine-conjugated phalloidin staining of F-actin in sh-NC/sh1-CHRDL1 CAL27 cells, examined using confocal microscopy. The arrows indicate filopodia. (B, C) The expression of EMT markers (Fibronectin, N-cadherin, E-cadherin, and Cytokeratin 18) in CAL27 cells after lentivirus transfection was detected by Western blot and quantitatively analyzed. All experiments were performed three times, and one representative experiment is shown. Error bars represent the standard deviation. ** p < 0.01, *** p < 0.001.

Article Snippet: Tissue sections were further incubated with primary antibody against CHRDL1 (1:150 dilution, Zen-Bio) at 4°C for 12 h and then washed and incubated with secondary antibody (1:100 dilution, ZSJB-Bio) at 37°C for 1 h. Finally, slides were incubated with diaminobenzidine (DAB, ZSJB-Bio) and counterstained with hematoxylin.

Techniques: Fluorescence, Staining, Confocal Microscopy, Expressing, Transfection, Western Blot, Standard Deviation

Journal: Frontiers in Oncology

Article Title: Chordin-Like 1 Regulates Epithelial-to-Mesenchymal Transition and Metastasis via the MAPK Signaling Pathway in Oral Squamous Cell Carcinoma

doi: 10.3389/fonc.2022.862751

Figure Lengend Snippet: CHRDL1 regulates the MAPK signaling pathway in CAL27 cells. (A) Heatmap of differentially expressed genes between OV-CHRDL1 and OV-NC (left) and between sh1-CHRDL1 and sh-NC (right). Green indicates downregulated genes, and red indicates upregulated genes. The thresholds for up- and downregulation were a +2.0-fold and -2.0-fold change, respectively, and a p adj < 0.05. (B) Top 10 GO-BP terms enriched among DEGs from OV-CHRDL1 CAL27 cells and sh1-CHRDL1 CAL27 cells. (C) Top 10 KEGG pathways enriched among DEGs from OV-CHRDL1 CAL27 cells and sh1-CHRDL1 CAL27 cells. (D, E) Protein expression of MAPK signaling pathway-related markers in CAL27 cells after lentivirus transfection. Error bars represent the standard deviation. ** p < 0.01, *** p < 0.001.

Article Snippet: Tissue sections were further incubated with primary antibody against CHRDL1 (1:150 dilution, Zen-Bio) at 4°C for 12 h and then washed and incubated with secondary antibody (1:100 dilution, ZSJB-Bio) at 37°C for 1 h. Finally, slides were incubated with diaminobenzidine (DAB, ZSJB-Bio) and counterstained with hematoxylin.

Techniques: Expressing, Transfection, Standard Deviation

Journal: Frontiers in Oncology

Article Title: Chordin-Like 1 Regulates Epithelial-to-Mesenchymal Transition and Metastasis via the MAPK Signaling Pathway in Oral Squamous Cell Carcinoma

doi: 10.3389/fonc.2022.862751

Figure Lengend Snippet: Effects of CHRDL1 on CAL27 cells in vivo . (A) Representative images of the lungs of BALB/c nude mice at 9 weeks after inoculation with OV-NC/OV-CHRDL1 (left) or sh-NC/sh1-CHRDL1 (right) CAL27 cells by tail vein injection. The red arrows show nodules on the lung surfaces. (B) Weights of the lungs of BALB/c-nude mice inoculated with OV-NC/OV-CHRDL1 (left) or sh-NC/sh1-CHRDL1 (right) CAL27 cells by tail vein injection. (C) Photomicrographs of hematoxylin-and-eosin-stained lungs of mice injected with OV-NC/OV-CHRDL1 (up) or sh-NC/sh1-CHRDL1 (down) CAL27 cells. Error bars represent the standard deviation. * p < 0.05, *** p < 0.001, **** p < 0.0001.

Article Snippet: Tissue sections were further incubated with primary antibody against CHRDL1 (1:150 dilution, Zen-Bio) at 4°C for 12 h and then washed and incubated with secondary antibody (1:100 dilution, ZSJB-Bio) at 37°C for 1 h. Finally, slides were incubated with diaminobenzidine (DAB, ZSJB-Bio) and counterstained with hematoxylin.

Techniques: In Vivo, Injection, Staining, Standard Deviation