nid2 polyclonal antibody  (Proteintech)

Journal: Molecular Human Reproduction

Article Title: Upregulated TIMP1 facilitates and coordinates myometrial contraction by decreasing collagens and cell adhesive capacity during human labor

doi: 10.1093/molehr/gaad034

Figure Lengend Snippet: Different cell adhesive properties in different human myometrial smooth muscle cells (hMSMCs). ( A ) Cell adhesive properties in nonpregnant, nonlabor and in-labor term hMSMCs. *IL versus NL, P < 0.05. ( B ) Cell adhesion properties in laboring hMSMCs with/without si-TIMP1 or oe-TIMP1. *si-TIMP1 versus the control, P < 0.05. ( C ) Representative western blotting of SPP1 (70 kDa), NID2 (151 kDa), and VCAN (373 kDa) in laboring hMSMCs with/without si-TIMP1 knockdown. See for full western blots. ( D ) Ratio of SPP1, NID2, and VCAN/β-actin expression in western blotting. ( E ) RNA-seq analysis shows the mRNA expression before and after si-TIMP1 knockdown. TIMP1, tissue inhibitor of metalloproteinase 1; SPP1, osteopontin; NID2, nidogen-2; VCAN, versican; NL, nonlabor; IL, in-labor. Data are presented as mean ± SEM from 3 to 10 independent experiments. * P < 0.05.

Article Snippet: The antibodies used were rabbit polyclonal β-actin antibody (1:5000, ab8226, Abcam, Cambridge, UK), rabbit monoclonal TIMP1 antibody (1:1000, ab211926, Abcam, Cambridge, UK), rabbit polyclonal NID2 (1:1000, DF9704, Affinity Biosciences, Changzhou, China), rabbit monoclonal VCAN (1:1000, ab270444, Abcam, Cambridge, UK), rabbit polyclonal SPP1 (1:1000, GB112328, Servicebio, Wuhan, China), and rabbit polyclonal GAPDH (1:1000, AF7021, Affinity Biosciences, Changzhou, China).

Techniques: Adhesive, Control, Western Blot, Knockdown, Expressing, RNA Sequencing

Journal: Oncotarget

Article Title: Metastasis-suppressing NID2 , an epigenetically-silenced gene, in the pathogenesis of nasopharyngeal carcinoma and esophageal squamous cell carcinoma

doi: 10.18632/oncotarget.12889

Figure Lengend Snippet: ( A ) The average methylation level of NID2 derived from our previous methylome data in NPC and ESCC. The vertical broken line shows the region covering the promoter CpG island (chr14: 52534582–52536722) and the bottom figures show a close-up view of changes in methylation. Methylation level is presented as β value (β = M/(U+M+100), M: signal intensity of the methylated allele, U: signal intensity of the unmethylated allele). The y-axis shows the average methylation level in tumors (orange line) and non-cancer controls (blue line), respectively. Within this region, the methylation levels in multiple CpG sites of both NPC and ESCC patients are consistently higher than those of non-cancer controls, with adjusted p value < 0.05 estimated by LIMMA analysis using the transformed β values as previously described . Significance level of each selected probes were shown in . ( B ) MMCT of chromosome 14 was previously performed using HONE1 as the recipient cell line . qPCR analysis of tumor-suppressive microcell hybrids (MCHs) and their tumor segregant (TS) cell lines, which are no longer tumor-suppressive, showed that NID2 expression was down-regulated in all five TS cell lines, when compared to their respective MCHs. Asterisk (*) indicates samples with more than two-fold differences compared to its MCHs.

Article Snippet: IHC staining was performed on NPC biopsies using anti-NID2 (SAB1409003) polyclonal antibody from Sigma-Aldrich (St Louis, MO, USA), following procedures as previously reported [ ].

Techniques: Methylation, Derivative Assay, Transformation Assay, Expressing

Journal: Oncotarget

Article Title: Metastasis-suppressing NID2 , an epigenetically-silenced gene, in the pathogenesis of nasopharyngeal carcinoma and esophageal squamous cell carcinoma

doi: 10.18632/oncotarget.12889

Figure Lengend Snippet: ( A ) In 37 of 50 (74.0%, p -value = 0.0001) NPC biopsies, the NID2 promoter hypermethylation was observed. The NID2 promoter was methylated in 12 of 15 (80.0%, p -value = 0.1281) ESCC biopsies, as shown by MS-HRM results. ( B ) qPCR analysis showed that expression of NID2 is down-regulated in 50% (37/74) of NPC biopsies and 43% (12/28) of ESCC biopsies showing more than two-fold down-regulation of NID2 expression. ( C ) All 7 NPC cell lines showed down-regulation of NID2 expression, after normalization to the immortalized nasopharyngeal cell line. 75% of the ESCC cell lines (12 of 16) have decreased NID2 expression, when compared to the immortalized normal esophageal epithelial cell line. The housekeeping gene GAPDH was used as an internal control for all qPCR analysis and the asterisks (*) indicate samples with more than two-fold up-regulation (> 2.0) or down-regulation (< 0.5).

Article Snippet: IHC staining was performed on NPC biopsies using anti-NID2 (SAB1409003) polyclonal antibody from Sigma-Aldrich (St Louis, MO, USA), following procedures as previously reported [ ].

Techniques: Methylation, Expressing

Journal: Oncotarget

Article Title: Metastasis-suppressing NID2 , an epigenetically-silenced gene, in the pathogenesis of nasopharyngeal carcinoma and esophageal squamous cell carcinoma

doi: 10.18632/oncotarget.12889

Figure Lengend Snippet: ( A ) Western blot analysis confirmed re-expression of NID2 in selected NPC (HONE1) and ESCC (KYSE30) cell lines, which have down-regulated NID2 expression. Over-expression of NID2 was detected in cell lysates, as well as being secreted into the conditioned media. Coomassie blue staining of total protein was used as a loading control for the conditioned media. ( B ) 2D colony formation assays of HONE1 and KYSE30 cells. Re-expression of NID2 significantly reduced their colony-forming abilities to around 40% (** p < 0.01). Data shown are the mean from three independent experiments ± S.D. ( C ) In 3D Matrigel culture, cells re-expressing NID2 resulted in significantly lower numbers of tumor spheres (≥ 100 μm), when compared to the VA in both NID2-expressing HONE1 and KYSE30 cells (** p < 0.01). Data shown are the mean from three independent experiments ± S.D.

Article Snippet: IHC staining was performed on NPC biopsies using anti-NID2 (SAB1409003) polyclonal antibody from Sigma-Aldrich (St Louis, MO, USA), following procedures as previously reported [ ].

Techniques: Western Blot, Expressing, Over Expression, Staining

Journal: Oncotarget

Article Title: Metastasis-suppressing NID2 , an epigenetically-silenced gene, in the pathogenesis of nasopharyngeal carcinoma and esophageal squamous cell carcinoma

doi: 10.18632/oncotarget.12889

Figure Lengend Snippet: ( A ) Cell migration ability was evaluated by the wound healing assay. Microscopic images were taken at 0th hour and 24th hour to monitor the extent of wound closure. Both NID2-expressing HONE1 and KYSE30 cells showed impaired wound healing ability, when compared to the VA controls (* p < 0.05). ( B ) Representative fields of migrated or invaded cells at 100x magnification. Re-expression of NID2 resulted in significant reduction of migration and invasion abilities of HONE1 and KYSE30 cells (* p < 0.05, ** p < 0.01). Data shown are the mean from three independent experiments ± S.D.

Article Snippet: IHC staining was performed on NPC biopsies using anti-NID2 (SAB1409003) polyclonal antibody from Sigma-Aldrich (St Louis, MO, USA), following procedures as previously reported [ ].

Techniques: Migration, Wound Healing Assay, Expressing

Journal: Oncotarget

Article Title: Metastasis-suppressing NID2 , an epigenetically-silenced gene, in the pathogenesis of nasopharyngeal carcinoma and esophageal squamous cell carcinoma

doi: 10.18632/oncotarget.12889

Figure Lengend Snippet: ( A ) Phosphorylation study of NID2-expressing cells using the Human Phospho-Kinase Antibody Array. Reduced phosphorylation of (1) FAK (Y397), (3) STAT1 (Y701), (4) Akt (T308), (5) STAT4 (Y693), (6) RSK1/2/3 (S380/S386/S377), and (7) PLCγ (Y783), and increased phosphorylation of (2) ERK1/2 (T202/Y204, T185/Y187) in the NID2-expressing cells was observed. The bar chart shows the relative quantification of the array data, normalized to VA control (* p < 0.05). Blue arrows indicated significant changes that were further validated using Western blot. ( B ) Re-expression of NID2 in HONE1 and KYSE30 cell lines resulted in reduced phosphorylation levels of FAK (Y397/Y925) and PLCγ (Y783), as shown by representative Western blot images. ( C ) The levels of phosphorylated EGFR at five tyrosine sites (Y992/Y1068/Y1086/Y1148/Y1173) were decreased by the re-expression of NID2 in both HONE1 and KYSE30. Downstream Akt pathways were also suppressed by NID2, as shown by the reduced phosphorylation level of Akt (T308/S473). Numbers at the bottom of each band represent the relative quantification of the protein level, normalized to the VA control.

Article Snippet: IHC staining was performed on NPC biopsies using anti-NID2 (SAB1409003) polyclonal antibody from Sigma-Aldrich (St Louis, MO, USA), following procedures as previously reported [ ].

Techniques: Expressing, Ab Array, Western Blot

Journal: Oncotarget

Article Title: Metastasis-suppressing NID2 , an epigenetically-silenced gene, in the pathogenesis of nasopharyngeal carcinoma and esophageal squamous cell carcinoma

doi: 10.18632/oncotarget.12889

Figure Lengend Snippet: ( A ) Representative images of nude mouse bioluminescent imaging to monitor the extent of in vivo metastasis after intrasplenic injection ( B ) Lesions on liver were apparent to the naked eye, for those in which metastasis occurred, and are indicated with arrows. None of the excised livers in the NID2 group showed overt tumor lesions. ( C ) Comparison of in vivo metastasis ability between the two groups revealed a significantly strong suppression of liver metastasis in the NID2 group, in which 0 of 12 mice developed tumors, while in contrast, 10 of 11 mice in the vector control group developed liver metastasis (** p < 0.0001) ( D ) Histological examination of the excised livers confirms the presence of multiple tumors. Representative H&E stained images of mouse liver from the VA group with metastatic undifferentiated carcinoma and mouse liver from NID2-expressing group, with no observable tumor.

Article Snippet: IHC staining was performed on NPC biopsies using anti-NID2 (SAB1409003) polyclonal antibody from Sigma-Aldrich (St Louis, MO, USA), following procedures as previously reported [ ].

Techniques: Imaging, In Vivo, Injection, Plasmid Preparation, Staining, Expressing

Journal: Oncotarget

Article Title: Metastasis-suppressing NID2 , an epigenetically-silenced gene, in the pathogenesis of nasopharyngeal carcinoma and esophageal squamous cell carcinoma

doi: 10.18632/oncotarget.12889

Figure Lengend Snippet: ( A ) In vitro cell proliferation was assessed using the MTT assay. Re-expression of NID2 did not affect HONE1 and KYSE30 cell growth. ( B ) Nude mouse subcutaneous injection was used to assess the effect of NID2 on in vivo tumor growth. Using both HONE1 and KYSE30, there were no significant differences in the tumor growth kinetics between the NID2-expressing and VA groups. Each data point was plotted as an average tumor volume of six tumor sites ± S.E.M.

Article Snippet: IHC staining was performed on NPC biopsies using anti-NID2 (SAB1409003) polyclonal antibody from Sigma-Aldrich (St Louis, MO, USA), following procedures as previously reported [ ].

Techniques: In Vitro, MTT Assay, Expressing, Injection, In Vivo