recombinant mouse b7h3 protein  (MedChemExpress)

Journal: Cancer Genomics & Proteomics

Article Title: ATIC Knockdown Reduces B7-H3 Expression and Oncogenic Signaling in Upper Tract Urothelial Carcinoma Cells

doi: 10.21873/cgp.20575

Figure Lengend Snippet: Identification of downstream signaling pathways of ATIC using isobaric labeling with tandem mass tags and liquid chromatography tandem mass spectrometry proteomics analysis in BFTC909 cells transfected with negative control siRNA or ATIC siRNA. (A) In the volcano plot, scattered points represent the differential fold changes of various proteins. The x-axis shows the log2 fold change, while the y-axis displays the −log10-adjusted p-Values. Blue indicates a significant decrease, and red indicates a significant increase (p<0.05). (B) Differentially expressed protein (DEP) rank plot depicting the fold change between samples against the rank of the protein expression value. The highest fold change corresponds to rank 1 (left x-axis), while the lowest corresponds to the last rank (right x-axis). (C) Western blot analysis showed that the knockdown of ATIC reduced B7-H3, prion protein, RAC2, and NT5E protein expression in BFTC909 cells. (D) Quantification of the protein levels of ATIC, B7-H3, prion protein, RAC2, and NT5E from (C) (n=3). Results are shown as the mean±standard deviation (SD); *p<0.05 and **p<0.01 using two‐tailed t‐test for (D).

Article Snippet: Membranes were incubated overnight at 4°C with anti-ATIC (MA1-086, Invitrogen, Waltham, MA, USA), β-actin (#3700, Cell Signaling Technology, Danvers, MA, USA), α-tubulin (NB100-690, Novus Biologicals, Centennial, CO, USA), B7-H3 (#14058, Cell Signaling Technology), Prion Protein (A18058, ABclonalbio, New Taipei, Taiwan, ROC), RAC2 (A1139, ABclonalbio), NT5E (A25914, ABclonalbio), Fibronectin 1 (#26836, Cell Signaling Technology), Slug (NBP2-52570, Novus), Cyclin A2 (#4656, Cell Signaling Technology), Cyclin B1 (#4138, Cell Signaling Technology), p57 (NBP1-89917, Novus), phospho-mTOR (Ser2448) (SAB4504476, Sigma-Aldrich), mTOR (#2983, Cell Signaling Technology), phospho-AKT (Thr308) (#9275, Cell Signaling Technology), AKT (#9272, Cell Signaling Technology), phospho-p38 MAPK (Thr180/Tyr182) (#9211, Cell Signaling Technology), p38 MAPK (#9212, Cell Signaling Technology), phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (#9101, Cell Signaling Technology), and p44/42 MAPK (Erk1/2) (#9102, Cell Signaling Technology).

Techniques: Protein-Protein interactions, Labeling, Liquid Chromatography, Mass Spectrometry, Transfection, Negative Control, Expressing, Western Blot, Knockdown, Standard Deviation, Two Tailed Test

Journal: Cancer Genomics & Proteomics

Article Title: ATIC Knockdown Reduces B7-H3 Expression and Oncogenic Signaling in Upper Tract Urothelial Carcinoma Cells

doi: 10.21873/cgp.20575

Figure Lengend Snippet: Functional characterization of ATIC downstream targets in BFTC909 cells. (A, B) Western blot validation of siRNA-mediated knockdown efficiency for B7-H3, prion protein, RAC2, and NT5E in BFTC909 cells (n=3). (C) Silencing of B7-H3, RAC2, or NT5E significantly reduced cell proliferation, as assessed using cell viability assays (n=4). (D, E) Knockdown of B7-H3, prion protein, RAC2, or NT5E markedly suppressed BFTC909 cell migration and invasion (n=3). (F) Knockdown of B7-H3 decreased fibronectin 1, slug, cyclin A2, and cyclin B1 expression while increasing p57 levels (n=3). (G) Silencing of prion protein reduced fibronectin 1 and cyclin A2 expression while upregulating p57 expression (n=3). (H) Suppression of RAC2 diminished fibronectin 1 and slug expression with a concomitant increase in p57 levels (n=3). (I) Depletion of NT5E decreased fibronectin 1 and elevated p57 expression levels (n=3). The results are shown as the mean±standard deviation (SD); *p<0.05, **p<0.01, and ***p<0.001; NS: Nonsignificant using two‐tailed t‐test for (B), (F), (G), (H) and (I), and ANOVA followed by Fisher’s LSD test for (C), (D) and (E).

Article Snippet: Membranes were incubated overnight at 4°C with anti-ATIC (MA1-086, Invitrogen, Waltham, MA, USA), β-actin (#3700, Cell Signaling Technology, Danvers, MA, USA), α-tubulin (NB100-690, Novus Biologicals, Centennial, CO, USA), B7-H3 (#14058, Cell Signaling Technology), Prion Protein (A18058, ABclonalbio, New Taipei, Taiwan, ROC), RAC2 (A1139, ABclonalbio), NT5E (A25914, ABclonalbio), Fibronectin 1 (#26836, Cell Signaling Technology), Slug (NBP2-52570, Novus), Cyclin A2 (#4656, Cell Signaling Technology), Cyclin B1 (#4138, Cell Signaling Technology), p57 (NBP1-89917, Novus), phospho-mTOR (Ser2448) (SAB4504476, Sigma-Aldrich), mTOR (#2983, Cell Signaling Technology), phospho-AKT (Thr308) (#9275, Cell Signaling Technology), AKT (#9272, Cell Signaling Technology), phospho-p38 MAPK (Thr180/Tyr182) (#9211, Cell Signaling Technology), p38 MAPK (#9212, Cell Signaling Technology), phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (#9101, Cell Signaling Technology), and p44/42 MAPK (Erk1/2) (#9102, Cell Signaling Technology).

Techniques: Functional Assay, Western Blot, Biomarker Discovery, Knockdown, Migration, Expressing, Standard Deviation, Two Tailed Test

Journal: Cancer Genomics & Proteomics

Article Title: ATIC Knockdown Reduces B7-H3 Expression and Oncogenic Signaling in Upper Tract Urothelial Carcinoma Cells

doi: 10.21873/cgp.20575

Figure Lengend Snippet: Interactions among ATIC-associated downstream molecules in UTUC cells. (A) Western blot analysis showed that the knockdown of B7-H3 markedly reduced prion protein and RAC2 protein expression in BFTC909 cells (n=3). (B) Silencing of prion protein decreased RAC2 protein expression in BFTC909 cells (n=3). (C) Knockdown of RAC2 did not significantly alter NT5E protein levels in BFTC909 cells (n=3). The results are shown as the mean±standard deviation (SD); **p<0.01, ***p<0.001, NS: Nonsignificant, using two-tailed t-test.

Article Snippet: Membranes were incubated overnight at 4°C with anti-ATIC (MA1-086, Invitrogen, Waltham, MA, USA), β-actin (#3700, Cell Signaling Technology, Danvers, MA, USA), α-tubulin (NB100-690, Novus Biologicals, Centennial, CO, USA), B7-H3 (#14058, Cell Signaling Technology), Prion Protein (A18058, ABclonalbio, New Taipei, Taiwan, ROC), RAC2 (A1139, ABclonalbio), NT5E (A25914, ABclonalbio), Fibronectin 1 (#26836, Cell Signaling Technology), Slug (NBP2-52570, Novus), Cyclin A2 (#4656, Cell Signaling Technology), Cyclin B1 (#4138, Cell Signaling Technology), p57 (NBP1-89917, Novus), phospho-mTOR (Ser2448) (SAB4504476, Sigma-Aldrich), mTOR (#2983, Cell Signaling Technology), phospho-AKT (Thr308) (#9275, Cell Signaling Technology), AKT (#9272, Cell Signaling Technology), phospho-p38 MAPK (Thr180/Tyr182) (#9211, Cell Signaling Technology), p38 MAPK (#9212, Cell Signaling Technology), phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (#9101, Cell Signaling Technology), and p44/42 MAPK (Erk1/2) (#9102, Cell Signaling Technology).

Techniques: Western Blot, Knockdown, Expressing, Standard Deviation, Two Tailed Test

Journal: Cancer Genomics & Proteomics

Article Title: ATIC Knockdown Reduces B7-H3 Expression and Oncogenic Signaling in Upper Tract Urothelial Carcinoma Cells

doi: 10.21873/cgp.20575

Figure Lengend Snippet: The ATIC/B7-H3 axis modulates mTOR, AKT, ERK, and p38 signaling in BFTC909 cells. (A) Western blot analysis showing that ATIC knockdown reduces the phosphorylation of mTOR, AKT, ERK, and p38, and decreases total mTOR expression in BFTC909 cells (n=3). (B) Silencing of B7-H3 reduced total and phosphorylated mTOR levels, and decreased the phosphorylation of AKT, ERK, and p38 (n=3). (C) Prion protein knockdown decreased total and phosphorylated mTOR levels, and reduced the activation of AKT, ERK, and p38 (n=3). (D) RAC2 silencing primarily reduced total and phosphorylated mTOR expression. (E) NT5E knockdown had no significant effect on mTOR, AKT, ERK, or p38 activity (n=3). The results are shown as the mean±standard deviation (SD); *p<0.05, **p<0.01, ***p<0.001, NS: Nonsignificant, using two-tailed t-test.

Article Snippet: Membranes were incubated overnight at 4°C with anti-ATIC (MA1-086, Invitrogen, Waltham, MA, USA), β-actin (#3700, Cell Signaling Technology, Danvers, MA, USA), α-tubulin (NB100-690, Novus Biologicals, Centennial, CO, USA), B7-H3 (#14058, Cell Signaling Technology), Prion Protein (A18058, ABclonalbio, New Taipei, Taiwan, ROC), RAC2 (A1139, ABclonalbio), NT5E (A25914, ABclonalbio), Fibronectin 1 (#26836, Cell Signaling Technology), Slug (NBP2-52570, Novus), Cyclin A2 (#4656, Cell Signaling Technology), Cyclin B1 (#4138, Cell Signaling Technology), p57 (NBP1-89917, Novus), phospho-mTOR (Ser2448) (SAB4504476, Sigma-Aldrich), mTOR (#2983, Cell Signaling Technology), phospho-AKT (Thr308) (#9275, Cell Signaling Technology), AKT (#9272, Cell Signaling Technology), phospho-p38 MAPK (Thr180/Tyr182) (#9211, Cell Signaling Technology), p38 MAPK (#9212, Cell Signaling Technology), phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (#9101, Cell Signaling Technology), and p44/42 MAPK (Erk1/2) (#9102, Cell Signaling Technology).

Techniques: Western Blot, Knockdown, Phospho-proteomics, Expressing, Activation Assay, Activity Assay, Standard Deviation, Two Tailed Test

Journal: Cancer Genomics & Proteomics

Article Title: ATIC Knockdown Reduces B7-H3 Expression and Oncogenic Signaling in Upper Tract Urothelial Carcinoma Cells

doi: 10.21873/cgp.20575

Figure Lengend Snippet: The ATIC/B7-H3 axis regulates cisplatin sensitivity in BFTC909 cells. (A) Western blot analysis showing the protein expression of ATIC, B7-H3, prion protein, RAC2, and NT5E in BFTC909 cells after 48 h of treatment with cisplatin (0–20 μM). (B) Cell viability assays showing that the knockdown of ATIC, B7-H3, prion protein, RAC2, or NT5E significantly enhanced cisplatin-induced cytotoxicity in BFTC909 cells after 48 h of treatment (n=3). The results are shown as the mean±standard deviation (SD); **p<0.01 (vs. si-Control non-treatment); #p<0.05 (vs. si-Control CDDP 20 μM); ##p<0.01 (vs. si-Control CDDP 20 μM) using ANOVA followed by Fisher’s LSD test.

Article Snippet: Membranes were incubated overnight at 4°C with anti-ATIC (MA1-086, Invitrogen, Waltham, MA, USA), β-actin (#3700, Cell Signaling Technology, Danvers, MA, USA), α-tubulin (NB100-690, Novus Biologicals, Centennial, CO, USA), B7-H3 (#14058, Cell Signaling Technology), Prion Protein (A18058, ABclonalbio, New Taipei, Taiwan, ROC), RAC2 (A1139, ABclonalbio), NT5E (A25914, ABclonalbio), Fibronectin 1 (#26836, Cell Signaling Technology), Slug (NBP2-52570, Novus), Cyclin A2 (#4656, Cell Signaling Technology), Cyclin B1 (#4138, Cell Signaling Technology), p57 (NBP1-89917, Novus), phospho-mTOR (Ser2448) (SAB4504476, Sigma-Aldrich), mTOR (#2983, Cell Signaling Technology), phospho-AKT (Thr308) (#9275, Cell Signaling Technology), AKT (#9272, Cell Signaling Technology), phospho-p38 MAPK (Thr180/Tyr182) (#9211, Cell Signaling Technology), p38 MAPK (#9212, Cell Signaling Technology), phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (#9101, Cell Signaling Technology), and p44/42 MAPK (Erk1/2) (#9102, Cell Signaling Technology).

Techniques: Western Blot, Expressing, Knockdown, Standard Deviation, Control