nid1  (Proteintech)

Journal: Journal of Translational Internal Medicine

Article Title: Development of a broadly potent neutralizing antibody targeting Nidogen 1 effectively inhibits cancer growth and metastasis in preclinical tumor models

doi: 10.1515/jtim-2025-0008

Figure Lengend Snippet: Upregulation of NID1 is associated with poor prognosis in HCC. A, Analysis of NID1 expression in tumor tissue (T, n = 374) and non-cancerous tissues (N, n = 49) using TCGA datasets of HCC. B, Kaplan-Meier curves comparing progression-free survival (PFS) of patients with high and low NID1 expressions in HCC stratified by median level, based on the Kaplan-Meier Plotter database ( http://kmplot.com/analysis ). C, Immunohistochemistry of NID1 was performed on TMA comprising paired T and NT tissues ( n = 80). The NID1 intensity score was analyzed. D, The pie chart illustrates the number of cases with overexpression, underexpression and no change in NID1 ( n = 80). E, Representative images of NID1 expression with intensity scores (0–3). Scale bar, 100 μm. F, Representative images of cases showing overexpression (T > NT), underexpression (T < NT) and no change (T = NT) in NID1. Scale bar, 100 μm. G, The overall IHC score of NID1 expression was plotted ( n = 80).

Article Snippet: For NID1 overexpression NID1 clone, the NID1 fragment (nucleotides 21-3123; Accession No. BC045606.1) was released from NID1/Entactin cDNA ORF Clone (Sino Biological) and subcloned into pMH-SFB cloning vector (Addgene) using gateway recombinational cloning (Invitrogen).

Techniques: Expressing, Immunohistochemistry, Over Expression

Journal: Journal of Translational Internal Medicine

Article Title: Development of a broadly potent neutralizing antibody targeting Nidogen 1 effectively inhibits cancer growth and metastasis in preclinical tumor models

doi: 10.1515/jtim-2025-0008

Figure Lengend Snippet: Generation of monoclonal anti-NID1 antibody. A, Schematic diagram illustrates the structural organization of NID1. The location of the 12-residue peptide for anti-NID1 antibody production is indicated by arrowhead. The location of epitope targets by the anti-NID1 antibody generated by the hybridoma clone is indicated by red arrowhead. B, The structural organization of NID1 and NID2. Homologous 12-residue peptide sequences in NID1 and NID2 are indicated by red arrowhead. Alignment of amino acid sequences of the selected epitope sequence between NID1 and NID2. Same residues between NID1 and NID2 are highlighted. C, Coomassie blue-stained polyacrylamide gel showing the purified anti-NID1 antibody obtained from hybridoma cells. D, Western blot analysis of NID1 expression in HLE cells stably transfected with backbone vector (XPack) and plasmid expressing NID1 (XP-NID1) using anti-NID1 antibody. E, Western blot analysis of NID1 expression in HCC cell lines using anti-NID1 antibody. F, Total cell lysates of MHCC97L cells was immunoprecipitated using anti-NID1 antibody. The immunoprecipitated proteins were subjected to immunoblotting using anti-NID1 antibody. G, Normal human hepatocyte MIHA cells were incubated with PBS or 5 μg/mL of anti-NID1 antibody. Number of cells was counted every 2 days. Data are represented as the mean ± SEM. NS, not significant from Student’s t -test.

Article Snippet: For NID1 overexpression NID1 clone, the NID1 fragment (nucleotides 21-3123; Accession No. BC045606.1) was released from NID1/Entactin cDNA ORF Clone (Sino Biological) and subcloned into pMH-SFB cloning vector (Addgene) using gateway recombinational cloning (Invitrogen).

Techniques: Residue, Generated, Sequencing, Staining, Purification, Western Blot, Expressing, Stable Transfection, Transfection, Plasmid Preparation, Immunoprecipitation, Incubation

Journal: Journal of Translational Internal Medicine

Article Title: Development of a broadly potent neutralizing antibody targeting Nidogen 1 effectively inhibits cancer growth and metastasis in preclinical tumor models

doi: 10.1515/jtim-2025-0008

Figure Lengend Snippet: Anti-NID1 antibody inhibits HCC growth and tumorigenesis. A, Immunoblots showing NID1 expression in the total cell lysate (TCL) of MHCC97 and LMHCCLM3 cells treated with anti-NID1 antibody. B, The colony forming ability of MHCC97L and MHCCLM3 cells treated with or without anti-NID1 antibody was assessed by colony formation assay. Representative images of colonies are shown (left). The number of colonies were counted and plotted (right). C, Schematic diagram of orthotopic liver implantation model. orthotopic liver implantation of luciferase-labelled MHCC97L tumor seed ( n = 7). Two weeks later, PBS, IgG and anti-NID1 antibody (400 μg/mouse) were injected intraperitoneally once a week for 21 days. Development of liver tumor was analyzed 3 weeks after liver implantation. D, At the end of experiment, tumors developed were excised and tumor size and weight were measured. Image showing the liver and tumors (left). Tumor weight (middle) and volume (right) were plotted. E, Body weight was measured regularly for 3 weeks. F, Immunohistochemistry of Ki67 staining of the excised tumors. The number of cells with positive Ki67 staining was counted. Analysis of the migration and invasiveness of MHCC97L (G) and MHCCLM3 (H) cells treated with or without anti-NID1 antibody using migration and invasion assays, respectively. Representative images of migrated and invaded cells are shown (left). Scale bar, 200 µm. Quantification of the number of migrated and invaded cells are plotted (right). I, Lung colonization of murine p53-/-; Myc hepatoblasts (1 × 105) after coinjection with anti-NID1 antibody (10 μg) via tail vein ( n = 5). Mice were subjected to bioluminescence imaging 14 days after injection. J, Bioluminescence imaging of mice at the end of the experiment. Quantification of the luciferase signal is shown. K, Bioluminescence imaging of dissected lung tissues. The intensity of luciferase signal is shown. L, Representative images of H & E staining of lung tissues. Insets show the enlarged area of the metastatic lesions. Scale bar, 150 µm. Data are represented as the mean ± SEM; * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001. NS, not significant from Student’s t -test. P < 0.05 is considered as statistically significant.

Article Snippet: For NID1 overexpression NID1 clone, the NID1 fragment (nucleotides 21-3123; Accession No. BC045606.1) was released from NID1/Entactin cDNA ORF Clone (Sino Biological) and subcloned into pMH-SFB cloning vector (Addgene) using gateway recombinational cloning (Invitrogen).

Techniques: Western Blot, Expressing, Colony Assay, Luciferase, Injection, Immunohistochemistry, Staining, Migration, Imaging

Journal: Journal of Translational Internal Medicine

Article Title: Development of a broadly potent neutralizing antibody targeting Nidogen 1 effectively inhibits cancer growth and metastasis in preclinical tumor models

doi: 10.1515/jtim-2025-0008

Figure Lengend Snippet: Anti-NID1 antibody effectively inhibits cell growth, migration and invasion of lung cancer, breast cancer and NPC in vitro. A, Kaplan-Meier analysis of overall survival of lung cancer and breast cancer patients with high and low NID1 expression based on the TCGA dataset. B, Immunoblots showing NID1 expression in HCC827, MDA-MB-231, and C666-1 cells treated with anti-NID1 antibody. Examination of the colony forming, migratory and invasive abilities of HCC827 (C), MDA-MB-231 (D), and C666-1 (E) cells treated with or without anti-NID1 antibody. Representative images of colonies, migrated and invaded cells are shown (left). Scale bar, 200 μm. Quantification of the number of colonies, migrated and invaded cells are plotted (right). Data are represented as the mean ± SEM; ** P < 0.01, *** P < 0.001, **** P < 0.0001. NS, not significant from Student’s t -test. P < 0.05 is considered as statistically significant.

Article Snippet: For NID1 overexpression NID1 clone, the NID1 fragment (nucleotides 21-3123; Accession No. BC045606.1) was released from NID1/Entactin cDNA ORF Clone (Sino Biological) and subcloned into pMH-SFB cloning vector (Addgene) using gateway recombinational cloning (Invitrogen).

Techniques: Migration, In Vitro, Expressing, Western Blot

Journal: Journal of Translational Internal Medicine

Article Title: Development of a broadly potent neutralizing antibody targeting Nidogen 1 effectively inhibits cancer growth and metastasis in preclinical tumor models

doi: 10.1515/jtim-2025-0008

Figure Lengend Snippet: Anti-NID1 antibody inhibits tumorigenesis of lung cancer, breast cancer and NPC in mouse models. HCC827 (A), MDA-MB-231 (D), and C666-1 (G) cells were injected subcutaneously into BALB/cAnN-nu mice. Administration of anti-NID1 antibody was started when the tumor volume reached 0.1 cm3. PBS, control IgG or anti-NID1 antibody (400 μg/mouse) were administered by intraperitoneal injection once every 3 days over a time period. B, E, F, Tumor size was measured regularly and plotted. C. F, I, At the end of experiment, tumors developed were excised and tumor size and weight were measured. Image showing the fixed excised tumors (left). Tumor weight (middle) and dimension (right) were plotted. J, Immunohistochemistry of Ki67 staining of the excised tumors. The number of Ki67 positively stained cells was analyzed. Scale bar, 100 μm. K, Body weight was measured regularly during the course of experiment. Data are represented as the mean ± SEM; * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001. NS, not significant from Student’s t -test. P < 0.05 is considered as statistically significant.

Article Snippet: For NID1 overexpression NID1 clone, the NID1 fragment (nucleotides 21-3123; Accession No. BC045606.1) was released from NID1/Entactin cDNA ORF Clone (Sino Biological) and subcloned into pMH-SFB cloning vector (Addgene) using gateway recombinational cloning (Invitrogen).

Techniques: Injection, Control, Immunohistochemistry, Staining

Journal: iScience

Article Title: Nidogen 1-enriched extracellular vesicles promote liver metastasis by inducing EMT and activating stellate cells

doi: 10.1016/j.isci.2025.113975

Figure Lengend Snippet: Extracellular vesicles from metastatic colorectal cancer cells (LM3) promote CRC tumor metastasis (A) Exosomes in the cell supernatants were identified by western blot experiments. (B) Exos in the cell supernatants were identified by transmission electron microscopy (upper) and NanoFCM (bottom). (C) Schematic diagram of the EV education mouse model. Nude mice were injected with EVs derived from SW480-P, SW480-LM3, km12-P, or km12-LM3 cells via the tail vein. Nude mice were injected with EVs derived from SW480-P, SW480-LM3, km12-P, or km12-LM3 cells via the tail vein once a week for 3 weeks (15 μg per week) prior to splenic injection of tumor seeds derived from SW480-P or km12-P cells ( n = 5). Analysis of liver metastases was performed 2 weeks after splenic injection. (D) In vivo fluorescence imaging and quantitative analysis of animals at the end of the experiment. (E) Venn diagram showing the overlap of upregulated genes identified via mRNA-seq in highly metastatic cells, serum exosome proteins in highly metastatic cells, and serum Exo proteins in colorectal cancer patients. (F) Representative IHC staining of NID1 expression in normal colon epithelium and paired primary CRC tissues. Scale bar: 50 μm. (G) NID1 expression was higher in CRC tissues ( n = 174) than in normal colon epithelium tissues ( n = 174) and higher in LM (+) CRC tissues ( n = 15) than in LM (−) CRC tissues ( n = 15) and in LM (−) CRC tissues ( n = 159). (H) Kaplan-Meier survival curves of overall survival in CRC patients with low ( n = 72) or high ( n = 102) NID1 expression. n.s. represents not significant; ∗∗ represents p < 0.01; ∗∗∗ represents p < 0.001, analyzed with paired t tests (D and G). Data are represented as mean ± SEM.

Article Snippet: NID1 , Proteintech , Cat. No. 13766-1-AP.

Techniques: Western Blot, Transmission Assay, Electron Microscopy, Injection, Derivative Assay, In Vivo, Fluorescence, Imaging, Immunohistochemistry, Expressing

Journal: iScience

Article Title: Nidogen 1-enriched extracellular vesicles promote liver metastasis by inducing EMT and activating stellate cells

doi: 10.1016/j.isci.2025.113975

Figure Lengend Snippet: The exosomal protein NID1 promotes liver metastasis of CRC tumors (A) EV mouse model comparing the effects of EVs from SW480/km12-LM3 and shNID1-1 SW480/km12-LM3 cells on CRC metastasis ( n = 5). (B and C) Image showing the luciferase signal of the animals at the end of the experiment. Quantification of the luciferase signal is shown. (D) Comparison of the effects of EVs from XPack and XP-NID1 HCT116 cells on CRC metastasis in an EV mouse model ( n = 5). (E and F) Image showing the luciferase signal of the animals at the end of the experiment. Quantification of the luciferase signal is shown. (G) Schematic diagram of the mouse model of in situ cecum injection combined with splenic injection. (H and I) In vivo imaging images of the mice and quantitative fluorescence statistics of the mouse liver region. n.s. represents not significant; ∗∗ represents p < 0.01; ∗∗∗ represents p < 0.001, analyzed with a t test (B, C, E, F, H, and I). Data are represented as mean ± SEM.

Article Snippet: NID1 , Proteintech , Cat. No. 13766-1-AP.

Techniques: Luciferase, Comparison, In Situ, Injection, In Vivo Imaging, Fluorescence

Journal: iScience

Article Title: Nidogen 1-enriched extracellular vesicles promote liver metastasis by inducing EMT and activating stellate cells

doi: 10.1016/j.isci.2025.113975

Figure Lengend Snippet: NID1 promotes colorectal cancer metastasis through the exosomal pathway (A–D) Transwell assays were performed to verify the effects of exosome-derived NID1 and the endogenous expression of NID1 on the invasive and metastatic ability of colorectal cancer cells in vitro . (E) Schematic diagram of a mouse spleen injected with tumor cells. (F) Splenic injection of SW480/km12 and NID1-overexpressing SW480/km12 cells in the presence/absence of cilengitide in vivo and NID1-overexpressing SW480/km12 cells overexpressing NID1 in vitro . (G) Quantification of the luciferase signal is shown. n.s. represents not significant; ∗∗∗ represents p < 0.001; ∗∗ represents p < 0.01, analyzed with paired t tests (A–D and G). Three independent experiments were performed. Data are represented as mean ± SEM.

Article Snippet: NID1 , Proteintech , Cat. No. 13766-1-AP.

Techniques: Derivative Assay, Expressing, In Vitro, Injection, In Vivo, Luciferase

Journal: iScience

Article Title: Nidogen 1-enriched extracellular vesicles promote liver metastasis by inducing EMT and activating stellate cells

doi: 10.1016/j.isci.2025.113975

Figure Lengend Snippet: The exosomal protein NID1 promotes epithelial mesenchymal transition in colorectal cancer cells (A) Western blotting was used to verify the effects of the EMT marker exosome-derived NID1 on colorectal cancer cells in vitro . (B) RT-qPCR was used to verify the effect of the EMT marker exosome-derived NID1 on colorectal cancer cells in vitro . (C) Western blotting was used to validate the expression of the EMT marker NID1 endogenously expressed in colorectal cancer cells in vitro . (D) RT-qPCR was used to verify the effect of the endogenous expression of the EMT marker NID1 on colorectal cancer cells in vitro . (E) Correlation of the expression of NID1 and EMT-associated mRNAs in primary tumors from the TCGA CRC patient cohort. (F) Relative mRNA expression (TPM) of EMT markers in LV5 and LV5-NID1 SW480 cells treated with GW4869 (10 μg mL-1) for 72 h. The TPM expression of EMT markers in LV5 and LV5-NID1 SW480 cells is shown in the following table. (G) KEGG pathway analysis revealed enrichment of genes associated with metastatic CRC cell lines. (H) Western blotting was used to verify the effect of the PI3K-Akt pathway marker exosome-derived NID1 on colorectal cancer cells in vitro . n.s. represents not significant; ∗∗ represents p < 0.01, analyzed with paired t test (B, D, and F). Three independent experiments were performed. Data are represented as mean ± SEM.

Article Snippet: NID1 , Proteintech , Cat. No. 13766-1-AP.

Techniques: Western Blot, Marker, Derivative Assay, In Vitro, Quantitative RT-PCR, Expressing

Journal: iScience

Article Title: Nidogen 1-enriched extracellular vesicles promote liver metastasis by inducing EMT and activating stellate cells

doi: 10.1016/j.isci.2025.113975

Figure Lengend Snippet: Ev-NID1 induces NET formation by facilitating IL-11 production in HSCs (A) Representative confocal microscope images showing the uptake of PKH26-labeled exosomes by LX-2 cells. Scale bar: 200 μm. (B) Bubble plot showing the GO signatures enriched in ev-NID1-stimulated LX-2 cells. (C) Immunofluorescence (IF) staining and quantification of NETs (H3Cit+ and MPO+ structures) in the liver metastases of mice treated with exosomes derived from ev-X-Pack HCT-116 and ev-XP-NID1 HCT116 cells for 12 h ( n = 18 Region of Metastatic Focus (RMFs) from 6 experimental replicates per group). Scale bar: 50 μm. (D) Cytokine array of the media of LX-2 cells pretreated with exosomes (X-Pack and XP-NID1) for 24 h. (E) ELISA results showing that IL-11 was upregulated in the supernatant of LX-2 cells stimulated with exosomes from XP-NID1. (F) Western blotting was used to verify the effect of the PI3K-Akt pathway marker exosome-derived NID1 on colorectal cancer cells in vitro . n.s. represents not significant; ∗∗∗ represents p < 0.001, analyzed with a t test (C and E). Three independent experiments were performed. Data are represented as mean ± SEM.

Article Snippet: NID1 , Proteintech , Cat. No. 13766-1-AP.

Techniques: Microscopy, Labeling, Immunofluorescence, Staining, Derivative Assay, Enzyme-linked Immunosorbent Assay, Western Blot, Marker, In Vitro

Journal: iScience

Article Title: Nidogen 1-enriched extracellular vesicles promote liver metastasis by inducing EMT and activating stellate cells

doi: 10.1016/j.isci.2025.113975

Figure Lengend Snippet: In liver metastases, NET formation is associated with IL-11, and targeting the IL-11 signaling pathway with a monoclonal antibody prevents CRLM (A) Immunofluorescence (IF) staining and quantification of IL-11+HSCs (IL-11+ and Desmin+) formed by LX-2 cells treated with exosomes derived from ev-X-Pack HCT-116 and ev-XP- NID1 HCT116 cells for 3 weeks. (B) Western blot analysis to characterize the expression of IL-11 in EV-NID1-treated LX-2 cells. (C) Representative confocal microscope images showing IL-11 expression in LX-2 cells treated with exosomes. Scale bar: 200 μm. (D) Western blot analysis of IL-11 expression in LX-2 cells under normal and quiescent conditions after treatment with EV-NID1 and the PI3K inhibitor (LY294002). (E) qPCR analysis of IL-11 expression in LX-2 cells under normal and quiescent conditions following treatment with EV-NID1 and the PI3K inhibitor (LY294002). (F) Splenic injection of XP-SW480/KM12 and XP-NID1 SW480/KM12 cells for liver metastasis experiments in the presence/absence of anti-IL-11. (G) Image showing the luciferase signal of the animals at the end of the experiment. Quantification of the luciferase signal is shown. (H) IF staining and quantification of NETs (H3Cit+ and MPO+ structures) in the liver metastases of mice treated with exosomes derived from ev-X-Pack HCT-116 and ev-XP-NID1 HCT116 cells for 12 h ( n = 18 RMFs from 6 experimental replicates per group). Scale bar: 50 μm. ∗∗ represents p < 0.01; ∗∗∗ represents p < 0.001, analyzed with a t test (A, E, and F). Three independent experiments were performed. Data are represented as mean ± SEM.

Article Snippet: NID1 , Proteintech , Cat. No. 13766-1-AP.

Techniques: Immunofluorescence, Staining, Derivative Assay, Western Blot, Expressing, Microscopy, Injection, Luciferase

Journal: Biology Direct

Article Title: NID1 promotes laryngeal cancer stemness via activating WNT pathway

doi: 10.1186/s13062-024-00548-0

Figure Lengend Snippet: NID1 is upregulated in LCA tissues, particularly in LCA tissues derived from relapsed patients. ( A ) NID1 was upregulated in LCA tissues, particularly in LCA tissues derived from relapsed patients. ( B ) Q-PCR and western blotting assays showed that NID1 was upregulated in LCA tissues (T) compared to the adjacent normal tissues (ANT). ( C ) Q-PCR and western blotting assays showed that NID1 was upregulated in the LCA cells. ( D ) Q-PCR and western blotting assays showed that NID1 was upregulated in LCA tissues derived from relapsed patients compared to the non- relapsed patients. Data represents the mean ± SD, * p < 0.05

Article Snippet: Antibodies that were used are the following: anti-NID1 antibody (MA5-23911, Thermo, Waltham, MA, USA), anti-cleaved Caspase-3 (ab2302, Abcam, Cambridge, UK), Cleaved PARP (ab32064, Abcam, Cambridge, UK), KLF4 (ab215036, Abcam, Cambridge, UK), OCT4 (ab181557, Abcam, Cambridge, UK), SOX2 (ab97959, Abcam, Cambridge, UK), NANOG (ab109250, Abcam, Cambridge, UK), β-Catenin (ab32572, Abcam, Cambridge, UK), EF1α (05-235, Merck, Darmstadt, Germany) and GAPDH (ab8245, Abcam, Cambridge, UK).

Techniques: Derivative Assay, Western Blot

Journal: Biology Direct

Article Title: NID1 promotes laryngeal cancer stemness via activating WNT pathway

doi: 10.1186/s13062-024-00548-0

Figure Lengend Snippet: NID1 is associated with poor outcome of LCA patients. ( A ) Kaplan-Meier analysis for overall survival, and recurrence free survival based on the expression of NID1. Data were come from TCGA, GSE27020 and GSE25727 databases. ( B ) Kaplan-Meier analysis for overall survival based on the expression of NID1. ( C ) The proportion of low NID1 or high NID1 level in LCA patients with or without Recurrence. Data represents the mean ± SD, *** p < 0.001

Article Snippet: Antibodies that were used are the following: anti-NID1 antibody (MA5-23911, Thermo, Waltham, MA, USA), anti-cleaved Caspase-3 (ab2302, Abcam, Cambridge, UK), Cleaved PARP (ab32064, Abcam, Cambridge, UK), KLF4 (ab215036, Abcam, Cambridge, UK), OCT4 (ab181557, Abcam, Cambridge, UK), SOX2 (ab97959, Abcam, Cambridge, UK), NANOG (ab109250, Abcam, Cambridge, UK), β-Catenin (ab32572, Abcam, Cambridge, UK), EF1α (05-235, Merck, Darmstadt, Germany) and GAPDH (ab8245, Abcam, Cambridge, UK).

Techniques: Expressing

Journal: Biology Direct

Article Title: NID1 promotes laryngeal cancer stemness via activating WNT pathway

doi: 10.1186/s13062-024-00548-0

Figure Lengend Snippet: NID1 promotes radiotherapy resistance in vitro. ( A ) Q-PCR and western blotting assays of the effect of NID1 shRNAs and overexpression vector. ( B ) Cell viability assay for the effect of NID1 knockdown or overexpression on radiotherapy. ( C ) Colony formation assay for the effect of NID1 knockdown or overexpression on radiotherapy. ( D ) Apoptosis assay for the effect of NID1 knockdown or overexpression on radiotherapy. Data represents the mean ± SD, * p < 0.05. ** p < 0.01

Article Snippet: Antibodies that were used are the following: anti-NID1 antibody (MA5-23911, Thermo, Waltham, MA, USA), anti-cleaved Caspase-3 (ab2302, Abcam, Cambridge, UK), Cleaved PARP (ab32064, Abcam, Cambridge, UK), KLF4 (ab215036, Abcam, Cambridge, UK), OCT4 (ab181557, Abcam, Cambridge, UK), SOX2 (ab97959, Abcam, Cambridge, UK), NANOG (ab109250, Abcam, Cambridge, UK), β-Catenin (ab32572, Abcam, Cambridge, UK), EF1α (05-235, Merck, Darmstadt, Germany) and GAPDH (ab8245, Abcam, Cambridge, UK).

Techniques: In Vitro, Western Blot, Over Expression, Plasmid Preparation, Viability Assay, Knockdown, Colony Assay, Apoptosis Assay

Journal: Biology Direct

Article Title: NID1 promotes laryngeal cancer stemness via activating WNT pathway

doi: 10.1186/s13062-024-00548-0

Figure Lengend Snippet: NID1 promotes radiotherapy resistance in vivo. ( A ) The schematic diagram for determining the effect of NID1 on radiotherapy using animal model. ( B ) The tumor sizes of xenograft tumor after radiotherapy treatment. ( C ) The tumor weights of xenograft tumor after radiotherapy treatment. ( D ) WB assay for the expression of Cleaved Caspase-3 and Cleaved RARP in tissues derived from xenograft tumor after radiotherapy treatment. ( E ) IHC staining assay for NID1 and β-Catenin in xenograft tumors. Scale bars, 60µM. Data represents the mean ± SD, ** p < 0.01

Article Snippet: Antibodies that were used are the following: anti-NID1 antibody (MA5-23911, Thermo, Waltham, MA, USA), anti-cleaved Caspase-3 (ab2302, Abcam, Cambridge, UK), Cleaved PARP (ab32064, Abcam, Cambridge, UK), KLF4 (ab215036, Abcam, Cambridge, UK), OCT4 (ab181557, Abcam, Cambridge, UK), SOX2 (ab97959, Abcam, Cambridge, UK), NANOG (ab109250, Abcam, Cambridge, UK), β-Catenin (ab32572, Abcam, Cambridge, UK), EF1α (05-235, Merck, Darmstadt, Germany) and GAPDH (ab8245, Abcam, Cambridge, UK).

Techniques: In Vivo, Animal Model, Expressing, Derivative Assay, Immunohistochemistry

Journal: Biology Direct

Article Title: NID1 promotes laryngeal cancer stemness via activating WNT pathway

doi: 10.1186/s13062-024-00548-0

Figure Lengend Snippet: NID1 promotes the self-renew of LCA stem cells. ( A ) Side populations assay for the effect of NID1 knockdown or overexpression on the self-renew of LCA stem cells. ( B ) Sphere formation assay for the effect of NID1 knockdown or overexpression on the self-renew of LCA stem cells. ( C ) WB assay for the expression of KLF4, OCT4, SOX2 and NANOG in LCA cells with NID1 knockdown or overexpression. ( D ) GSEA assay for the relation between NID1 expression and stem cell regulating genes. Data represents the mean ± SD, * p < 0.05. ** p < 0.01

Article Snippet: Antibodies that were used are the following: anti-NID1 antibody (MA5-23911, Thermo, Waltham, MA, USA), anti-cleaved Caspase-3 (ab2302, Abcam, Cambridge, UK), Cleaved PARP (ab32064, Abcam, Cambridge, UK), KLF4 (ab215036, Abcam, Cambridge, UK), OCT4 (ab181557, Abcam, Cambridge, UK), SOX2 (ab97959, Abcam, Cambridge, UK), NANOG (ab109250, Abcam, Cambridge, UK), β-Catenin (ab32572, Abcam, Cambridge, UK), EF1α (05-235, Merck, Darmstadt, Germany) and GAPDH (ab8245, Abcam, Cambridge, UK).

Techniques: Knockdown, Over Expression, Tube Formation Assay, Expressing

Journal: Biology Direct

Article Title: NID1 promotes laryngeal cancer stemness via activating WNT pathway

doi: 10.1186/s13062-024-00548-0

Figure Lengend Snippet: NID1 activates WNT pathway. ( A ) GSEA assay for the relation between NID1 expression and WNT pathway. ( B ) Luciferase reporter assay for the effect of NID1 on WNT pathway activation. ( C ) WB assay for the effect of NID1 on the nuclear location of β-Catenin in LCA cells. ( D ) IF assay for the effect of NID1 on the nuclear location of β-Catenin in LCA cells. ( E ) The effect of NID1 on the expression of the downstream genes of WNT pathway. Data represents the mean ± SD, ** p < 0.01

Article Snippet: Antibodies that were used are the following: anti-NID1 antibody (MA5-23911, Thermo, Waltham, MA, USA), anti-cleaved Caspase-3 (ab2302, Abcam, Cambridge, UK), Cleaved PARP (ab32064, Abcam, Cambridge, UK), KLF4 (ab215036, Abcam, Cambridge, UK), OCT4 (ab181557, Abcam, Cambridge, UK), SOX2 (ab97959, Abcam, Cambridge, UK), NANOG (ab109250, Abcam, Cambridge, UK), β-Catenin (ab32572, Abcam, Cambridge, UK), EF1α (05-235, Merck, Darmstadt, Germany) and GAPDH (ab8245, Abcam, Cambridge, UK).

Techniques: Expressing, Luciferase, Reporter Assay, Activation Assay

Journal: Biology Direct

Article Title: NID1 promotes laryngeal cancer stemness via activating WNT pathway

doi: 10.1186/s13062-024-00548-0

Figure Lengend Snippet: NID1 promotes LCA radiotherapy resistance and the self-renew of LCA stem cells via activating WNT pathway. ( A ) Colony formation assay for the effect of inhibition of WNT pathway on radiotherapy resistance in NID1-overexpressing LCA cells. ( B ) Apoptosis assay for the effect of inhibition of WNT pathway on radiotherapy resistance in NID1-overexpressing LCA cells. ( C ) Sphere formation assay for the effect of inhibition of WNT pathway on radiotherapy resistance in NID1-overexpressing LCA cells. ( D ) The correlation between NID1 expression of nuclear β-Catenin in LCA tissues determined by WB assay. Data represents the mean ± SD, ** p < 0.01

Article Snippet: Antibodies that were used are the following: anti-NID1 antibody (MA5-23911, Thermo, Waltham, MA, USA), anti-cleaved Caspase-3 (ab2302, Abcam, Cambridge, UK), Cleaved PARP (ab32064, Abcam, Cambridge, UK), KLF4 (ab215036, Abcam, Cambridge, UK), OCT4 (ab181557, Abcam, Cambridge, UK), SOX2 (ab97959, Abcam, Cambridge, UK), NANOG (ab109250, Abcam, Cambridge, UK), β-Catenin (ab32572, Abcam, Cambridge, UK), EF1α (05-235, Merck, Darmstadt, Germany) and GAPDH (ab8245, Abcam, Cambridge, UK).

Techniques: Colony Assay, Inhibition, Apoptosis Assay, Tube Formation Assay, Expressing