anti ptges3  (Novus Biologicals)

Journal: Biology Direct

Article Title: PTGES3 proteolysis using the liposomal peptide-PROTAC approach

doi: 10.1186/s13062-024-00580-0

Figure Lengend Snippet: PTGES3 was overexpressed and predicted poor prognosis in hepatocellular carcinoma (HCC). ( A ) Bioinformatics analysis of PTGES3 mRNA expression in multiple tumor samples by an online website GEPIA. ( B ) Protein expression analysis of PTGES3 in 165 HCC tumor samples and 165 normal tissues by UALCAN. ( C ) Kaplan-Meier survival analysis was used to measure the correlation between PTGES3 expression and overall survival in HCC from UALCAN website. ( D ) Immunohistochemical (IHC) staining pictures of PTGES3 protein in HCC and paracancerous samples from the Human Protein Atlas online database ( n = 3). ( E ) Representative IHC images of PTGES3 protein in paired HCC specimens and their normal liver tissues from the tissue microarray ( n = 90). ( F , G ) PTGES3 mRNA and protein expressions of HCC cell lines and normal liver epithelial THLE-2 cells from qRT-PCR ( F ) and western blotting ( G ) results ( n = 3)

Article Snippet: Ubiquitination level of PTGES3 was tested by CO-IP with anti-PTGES3 antibody (15216-1-AP, Proteintech Group).

Techniques: Expressing, Immunohistochemical staining, Immunohistochemistry, Microarray, Quantitative RT-PCR, Western Blot

Journal: Biology Direct

Article Title: PTGES3 proteolysis using the liposomal peptide-PROTAC approach

doi: 10.1186/s13062-024-00580-0

Figure Lengend Snippet: PTGES3 knockdown weakened hepatocellular carcinoma (HCC) cells proliferation and migration in vitro. ( A ) Western blotting was conducted to detect the capacity to manipulate PTGES3 expression by siRNAs ( n = 3). ( B , C ) EdU assays showed the growth of HCC cells ( n = 3). ( D , E ) CCK-8 experiments revealed the proliferative ability of HCC cells ( n = 3). ( F , G ) HCC cells were subjected to colony formation assays ( n = 3). ( H , I ) Transwell showed that the migrated ability of HCC cells ( n = 3). ( J , K ) Wound healing assays were performed to test the migration of HCC cells ( n = 3)

Article Snippet: Ubiquitination level of PTGES3 was tested by CO-IP with anti-PTGES3 antibody (15216-1-AP, Proteintech Group).

Techniques: Knockdown, Migration, In Vitro, Western Blot, Expressing, CCK-8 Assay

Journal: Biology Direct

Article Title: PTGES3 proteolysis using the liposomal peptide-PROTAC approach

doi: 10.1186/s13062-024-00580-0

Figure Lengend Snippet: The ablation of PTGES3 decreased the tumor growth in vivo. ( A ) Images of implanted tumors from nude mice injected with Hep3B and Huh7 cells subcutaneously ( n = 5). ( B ) Western blotting measured the PTGES3 expression in tumor samples ( n = 5). ( C ) Immunohistochemical (IHC) staining of PTGES3 protein in implanted tumors with stable transfection by shNC or shPTGES3 ( n = 5). ( D ) Tumor weight of nude mice ( n = 5). ( E ) Tumor volume of nude mice ( n = 5)

Article Snippet: Ubiquitination level of PTGES3 was tested by CO-IP with anti-PTGES3 antibody (15216-1-AP, Proteintech Group).

Techniques: In Vivo, Injection, Western Blot, Expressing, Immunohistochemical staining, Immunohistochemistry, Stable Transfection

Journal: Biology Direct

Article Title: PTGES3 proteolysis using the liposomal peptide-PROTAC approach

doi: 10.1186/s13062-024-00580-0

Figure Lengend Snippet: Identification of peptides targeting PTGES3 protein. ( A ) Name and sequence of peptides bound to PTGES3 protein. ( B ) Name and sequence of synthetic peptides conjugated with FITC and TAT sequence (GRKKRRQRRRPPQQ). ( C ) CCK-8 showed the growth rate of Huh7 cells treated with six synthetic peptides ( n = 3). ( D ) IF experiments showed the cellular localization of PTGES3 protein (red) and FITC labeled FIP-2 (green) in Huh7 cells ( n = 3). ( E ) The optimized binding modes between PTGES3 protein and F-2, Blue: PTGES3 protein; Green: F-2 peptide. ( F ) Temperature-dependent cellular thermal shift assay displayed the elevated thermal stability of PTGES3 in HCC cell lysates treated with FIP-2 ( n = 3). ( G ) Concentration-dependent cellular thermal shift experiment revealed the increased thermal stability of PTGES3 treated with FIP-2 in Hep3B and Huh7 cell lysates at 56 and 59°C, respectively ( n = 3)

Article Snippet: Ubiquitination level of PTGES3 was tested by CO-IP with anti-PTGES3 antibody (15216-1-AP, Proteintech Group).

Techniques: Sequencing, CCK-8 Assay, Labeling, Binding Assay, Thermal Shift Assay, Concentration Assay

Journal: Biology Direct

Article Title: PTGES3 proteolysis using the liposomal peptide-PROTAC approach

doi: 10.1186/s13062-024-00580-0

Figure Lengend Snippet: Preparation and characterization of PTGES3 liposome peptide-proteolysis targeting chimera (p-PROTAC). ( A ) Schematic diagram of the PTGES3 liposome p-PROTAC platform. ( B ) Particle size distribution of PTGES3 degrader liposomes collected via DLS. ( C ) Zeta potential of PTGES3 degrader liposomes ( n = 3). ( D ) Stability of PTGES3 degrader liposomes in vitro in presence of serum and various enzymes ( n = 3). ( E ) Stability of PTGES3 degrader liposomes Stored at 4 °C for 3 Months ( n = 3). ( F ) Transmission electron microscope picture of PTGES3 degrader liposomes ( n = 3)

Article Snippet: Ubiquitination level of PTGES3 was tested by CO-IP with anti-PTGES3 antibody (15216-1-AP, Proteintech Group).

Techniques: Liposomes, Zeta Potential Analyzer, In Vitro, Transmission Assay, Microscopy

Journal: Biology Direct

Article Title: PTGES3 proteolysis using the liposomal peptide-PROTAC approach

doi: 10.1186/s13062-024-00580-0

Figure Lengend Snippet: PTGES3-PROTAC induced PTGES3 proteolysis in a transcriptional independent manner. ( A ) Western blotting examination showed the PTGES3 degradation in HCC cells treated with different doses of FIP-2 or PTGES3 degrader liposomes ( n = 3). ( B ) Western blotting assays displayed the PTGES3 degradation in HCC cells treated with PTGES3-PROTAC at different time points ( n = 3). ( C ) Western blotting analysis of the ubiquitinated levels of PTGES3 in HCC cells treated with PTGES3-PROTAC and/or MG132 ( n = 3). ( D ) Western blotting analysis of the protein levels of PTGES3 in HCC cells treated with PTGES3-PROTAC and/or MG132 ( n = 3). ( E ) CCK-8 assays were performed to test proliferation of HCC cells treated with FIP-2 or PTGES3-PROTAC ( n = 3). ( F ) Western blotting assays showed the PTGES3 protein expression in HCC cells treated with FIP-2 or PTGES3-PROTAC ( n = 3). ( G ) Detection of PTGES3 mRNA expression by qRT-PCR in HCC cells treated with FIP-2 or PTGES3-PROTAC ( n = 3)

Article Snippet: Ubiquitination level of PTGES3 was tested by CO-IP with anti-PTGES3 antibody (15216-1-AP, Proteintech Group).

Techniques: Western Blot, Liposomes, CCK-8 Assay, Expressing, Quantitative RT-PCR

Journal: Biology Direct

Article Title: PTGES3 proteolysis using the liposomal peptide-PROTAC approach

doi: 10.1186/s13062-024-00580-0

Figure Lengend Snippet: PTGES3-PROTAC inhibited the malignant phenotype of HCC cells in vitro. ( A , B ) Representative and quantified results of the EdU incorporation assays. HCC cells were treated with FIP-2 or PTGES3-PROTAC ( n = 3). ( C , D ) The proliferation abilities were analyzed and quantified by colony formation assays. FIP-2 or PTGES3-PROTAC was used to treat HCC cells ( n = 3). ( E , F ) Transwell detection of HCC cells treated with FIP-2 or PTGES3-PROTAC. Representative photographs of transwell assays were presented ( E ). Statistical analysis of the transwell assays ( F , n = 3). ( G , H ) Representative images and quantified data of the wound healing assays. FIP-2 or PTGES3-PROTAC treating HCC showed the reduced migration ability ( n = 3)

Article Snippet: Ubiquitination level of PTGES3 was tested by CO-IP with anti-PTGES3 antibody (15216-1-AP, Proteintech Group).

Techniques: In Vitro, Migration

Journal: Biology Direct

Article Title: PTGES3 proteolysis using the liposomal peptide-PROTAC approach

doi: 10.1186/s13062-024-00580-0

Figure Lengend Snippet: PTGES3-PROTAC suppressed tumor growth in vivo. ( A ) Representative photographs of excised tumors from xenograft tumor models ( n = 5). ( B ) The protein levels of PTGES3 expression in subcutaneous xenografts from the control, FIP-2 treatment and PTGES3-PROTAC treatment groups ( n = 3). ( C ) Immunohistochemistry images of Ki67 and PTGES3 protein in xenograft tumors from the indicated groups ( n = 3). ( D ) Subcutaneous xenograft masses were excised and weighed at the endpoint ( n = 5). ( E ) Tumor growth curves of subcutaneous tumor formation in nude mice treated with FIP-2 or PTGES3-PROTAC ( n = 5). ( F ) H&E staining of major organ tissues harvested from tumor-bearing nude mice subcutaneously injected with Hep3B cells. The results showed no systemic toxicity ( n = 5). ( G ) Activities of serum alanine aminotransferase (ALT), aspartic acid transferase (AST), creatinine and blood urea nitrogen from nude mice subcutaneously injected with Hep3B cells ( n = 5)

Article Snippet: Ubiquitination level of PTGES3 was tested by CO-IP with anti-PTGES3 antibody (15216-1-AP, Proteintech Group).

Techniques: In Vivo, Expressing, Control, Immunohistochemistry, Staining, Injection