nod2 antibody (2d9) - bsa free  (Bio-Techne corporation)

Journal: International Journal of Oncology

Article Title: TRIM22 inhibits endometrial cancer progression through the NOD2/NF- k B signaling pathway and confers a favorable prognosis

doi: 10.3892/ijo.2020.5004

Figure Lengend Snippet: TRIM22 influences EnC cells by binding NOD2, and suppresses the migration, invasion and proliferation of EnC cells via the NOD2-NF- κb axis. (A) Immunofluorescence analysis of TRIM22 and NOD2 in normal endometrial tissues and endometrial tumor tissues from subjects undergoing hysterectomy and patients with EnC. TRIM22 and NOD2 colocalization was observed in the cytoplasm of both normal endometrial tissue and endometrial tumor tissue. Both TRIM22 and NOD2 colocalization and expression were decreased in EnC tissue. Scale bar, 20 µm. (B) Ishikawa cells were transfected with lentiviral construct encoding human TRIM22, and the interaction between TRIM22 and NOD2 was determined through co-immunoprecipitation examination. An empty vector construct was used as IP-negative controls. TRIM22 overexpression construct was examined. (C) Western blot analysis of IκBα degradation, phosphorylated I κb α (Ser 36), p65 and phosphorylated p65 (Ser 536) in Ishikawa TRIM22 OE cells. (D) Ishikawa cells were transfected with lentiviral construct encoding human TRIM22. Western blot analysis was applied to evaluate the location of p65 in Ishikawa and KLE cells. (E and F) Ishikawa cells were transfected with lentiviral construct encoding human TRIM22 at first and followed by transient transfection with shRNA-NF- κb -p65. Knockdown NF- κb -p65 attenuated the decrease in the migration, invasion and cell proliferation which was induced by the overexpression of TRIM22. The cell proliferative, migratory and invasive abilities of endometrial tumor cells increased. Scale bars: EdU, 50 µm; Transwell, 100 µm. ****P<0.0001. TRIM22, tripartite motif-containing 22; EnC, endometrial cancer; NOD2, nucleotide-binding oligomerization domain-containing protein 2.

Article Snippet: Primary antibodies were used at the following dilutions: TRIM22 antibody (1:350, NBP1-81795, Novus Biologicals, LLC), nucleotide binding oligomerization domain containing 2 (NOD2) antibody (1:250, NB100-524, Novus Biologicals, LLC), Ki-67 antibody (1:200, RB-9043-P1, eBioscience; Thermo Fisher Scientific, Inc.).

Techniques: Binding Assay, Migration, Immunofluorescence, Expressing, Transfection, Construct, Immunoprecipitation, Plasmid Preparation, Over Expression, Western Blot, shRNA, Knockdown

Journal: International Journal of Oncology

Article Title: TRIM22 inhibits endometrial cancer progression through the NOD2/NF- k B signaling pathway and confers a favorable prognosis

doi: 10.3892/ijo.2020.5004

Figure Lengend Snippet: TRIM22 influences EnC cells by binding NOD2, and suppresses the migration, invasion and proliferation of EnC cells via the NOD2-NF- κb axis. (A) Immunofluorescence analysis of TRIM22 and NOD2 in normal endometrial tissues and endometrial tumor tissues from subjects undergoing hysterectomy and patients with EnC. TRIM22 and NOD2 colocalization was observed in the cytoplasm of both normal endometrial tissue and endometrial tumor tissue. Both TRIM22 and NOD2 colocalization and expression were decreased in EnC tissue. Scale bar, 20 µm. (B) Ishikawa cells were transfected with lentiviral construct encoding human TRIM22, and the interaction between TRIM22 and NOD2 was determined through co-immunoprecipitation examination. An empty vector construct was used as IP-negative controls. TRIM22 overexpression construct was examined. (C) Western blot analysis of IκBα degradation, phosphorylated I κb α (Ser 36), p65 and phosphorylated p65 (Ser 536) in Ishikawa TRIM22 OE cells. (D) Ishikawa cells were transfected with lentiviral construct encoding human TRIM22. Western blot analysis was applied to evaluate the location of p65 in Ishikawa and KLE cells. (E and F) Ishikawa cells were transfected with lentiviral construct encoding human TRIM22 at first and followed by transient transfection with shRNA-NF- κb -p65. Knockdown NF- κb -p65 attenuated the decrease in the migration, invasion and cell proliferation which was induced by the overexpression of TRIM22. The cell proliferative, migratory and invasive abilities of endometrial tumor cells increased. Scale bars: EdU, 50 µm; Transwell, 100 µm. ****P<0.0001. TRIM22, tripartite motif-containing 22; EnC, endometrial cancer; NOD2, nucleotide-binding oligomerization domain-containing protein 2.

Article Snippet: Primary antibodies were used at the following dilutions: TRIM22 rabbit polyclonal antibody (1:200, NBP1-81795, Novus Biologicals, LLC), NOD2 antibody (1:1,000, NB100-524, Novus Biologicals, LLC), IκBα(E130) antibody (1:1,000, ab32518, Abcam), p-IκBα (Ser36) [EPR6235 ( )] antibody (1:10,000, ab133462, Abcam), NF-κ b p65 (E379) antibody (1:50,000, ab32536, Abcam), p-p65(Ser536) (EP2294Y) antibody (1:10,000, ab76302, Abcam), GAPDH (AG0766) antibody (1:10,000, 60004-1-Ig, Proteintech) and LaminB1 antibody (1:5,000, ab16048, Abcam) and secondary antibodies (rabbit, mouse, ZB-2301, ZB-2305, ZSGB-Bio) were used at 1:5,000.

Techniques: Binding Assay, Migration, Immunofluorescence, Expressing, Transfection, Construct, Immunoprecipitation, Plasmid Preparation, Over Expression, Western Blot, shRNA, Knockdown