Journal: Cancers

Article Title: Murlentamab, a Low Fucosylated Anti-Müllerian Hormone Type II Receptor (AMHRII) Antibody, Exhibits Anti-Tumor Activity through Tumor-Associated Macrophage Reprogrammation and T Cell Activation

doi: 10.3390/cancers13081845

Figure Lengend Snippet: Flow cytometry antibodies used.

Article Snippet: CD36-PEVio770 , Miltenyi , 130-110-742 , REA770 , .

Techniques: Flow Cytometry, In Vivo, In Vitro

Journal: Cancers

Article Title: Murlentamab, a Low Fucosylated Anti-Müllerian Hormone Type II Receptor (AMHRII) Antibody, Exhibits Anti-Tumor Activity through Tumor-Associated Macrophage Reprogrammation and T Cell Activation

doi: 10.3390/cancers13081845

Figure Lengend Snippet: Murlentamab opsonization of SKOV3-R2 + orients naïve macrophages and reprograms TAMs towards an M1-like profile. SKOV3-R2 + ovarian tumor cells were labeled with different 3C23K antibodies (3C23K-FcKO control, 3C23K-CHO normally fucosylated or murlentamab the low fucosylated form) and cultured in the presence of human monocyte-derived macrophages from healthy donors unstimulated (M0) or stimulated with M-CSF and IL-10 (TAMs). ( A ) The proportion of macrophages expressing M1/M2 membrane markers (CD32, CD64, CD80, TLR2, CD163, CD36 and CD206) was determined by flow cytometry after three days of co-culture with SKOV3-R2 + cells. ( B ) The release of cytokines (IL1β, IL12, TNFα, IL6, IFNγ, IL10) and chemokines (CCL2, CCL4, CCL5, CXCL9 and CXCL10) in the culture medium was determined by AlphaLISA after three days of co-culture with SKOV3-R2 + cells. Data shown (boxplots) are the results from three different experiments (performed with three different healthy donors). * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001. p values were determined using one-way ANOVA analysis followed by Tukey’s multiple comparisons test.

Article Snippet: CD36-PEVio770 , Miltenyi , 130-110-742 , REA770 , .

Techniques: Labeling, Cell Culture, Derivative Assay, Expressing, Membrane, Flow Cytometry, Co-Culture Assay

Journal: bioRxiv

Article Title: The atypical cadherin FAT1 is a novel regulator of STAT1, driving its pro-tumorigenic effect via the STAT1/PDCD4 axis in glioblastoma

doi: 10.1101/2023.09.22.558772

Figure Lengend Snippet: A : Scatter plot representing normalized mRNA expression of STAT1 and FAT1 in TCGA glioblastoma patient (cell 2013) available at GDC TCGA. STAT1 showed a positive Spearman correlation with FAT1 (correlation coefficient r = 0.18) in 141 patients. B: Scatter plot representing normalized mRNA expression of STAT4 and FAT1 in TCGA glioblastoma patient (cell 2013) available at GDC TCGA. STAT4 showed no significant Spearman correlation with FAT1. C, D: Dot plots representing normalized mRNA expression of FAT1 -STAT1 and FAT1-STAT4 respectively in microarray Loeffler Data available at R2 genomics and analysis platform of 70 patients. STAT1 showed a positive Spearman correlation with r = 0.24 and p value 0.04 while FAT1 and STAT4 have no significant correlation. E, F: Scatter plot representing the normalized mRNA expression accessed by Real-Time PCR of FAT1-STAT1 and FAT1-STAT4 respectively in 33 Glioblastoma resected samples. STAT1 showed a significant positive Spearman correlation of 0.53 with p value 0.0015 while STAT4 showed no positive correlation with FAT1.

Article Snippet: Antibodies used for Western blot were FAT1 antibody (NB100-2693; 1:500; 508 kDa), STAT1 antibody (9172S,1:1000), PDCD4 (D29C6,1:1000) (Cell Signaling technologies), β-Actin (AB8227; 1:5000; 42kDa).

Techniques: Expressing, Microarray, Real-time Polymerase Chain Reaction

Journal: bioRxiv

Article Title: The atypical cadherin FAT1 is a novel regulator of STAT1, driving its pro-tumorigenic effect via the STAT1/PDCD4 axis in glioblastoma

doi: 10.1101/2023.09.22.558772

Figure Lengend Snippet: A, B. Graphs representing fold mRNA expression in U87MG and LN229 cells treated with SiControl and siFAT1. Knockdown of FAT1 in U87MG cells and LN229 led to increased mRNA expression of PDCD4 (> 2 folds), STAT1 mRNA expression decreased (= 1.5 folds) assessed by qPCR experiment. C.D. Western Blot showing the effect of FAT1 knockdown on STAT1. E.F. Decrease in STAT1 Blot intensity (≥ 2folds) in U87MG and LN229 cells respectively, analyzed by Image Software. G. ICC Image of anti-STAT1 antibody showing STAT1 expression. H. Graph representing the mean fluorescence intensity of STAT1 antibody in control cells and FAT1 knockdown cells. Data presented as mean ±SD *p<0.05, **p<0.005 and ***p<0.0005.

Article Snippet: Antibodies used for Western blot were FAT1 antibody (NB100-2693; 1:500; 508 kDa), STAT1 antibody (9172S,1:1000), PDCD4 (D29C6,1:1000) (Cell Signaling technologies), β-Actin (AB8227; 1:5000; 42kDa).

Techniques: Expressing, Knockdown, Western Blot, Software, Fluorescence, Control

Journal: bioRxiv

Article Title: The atypical cadherin FAT1 is a novel regulator of STAT1, driving its pro-tumorigenic effect via the STAT1/PDCD4 axis in glioblastoma

doi: 10.1101/2023.09.22.558772

Figure Lengend Snippet: A, B. Increase in PDCD4 luciferase activity on FAT1 knockdown in LN229 cells (≥ 2 folds p≤ 0.005) and on STAT1 knockdown in LN229 cells (≥ 1.4 folds, p ≤ 0.05) C. Diagrammatic representation of the deleted nucleotides by SDM to disrupt STAT1 binding sites on PDCD4 promoter site 1 D. PDCD4 promoter luciferase activity increases by 2.8 folds (p-value: < 0.005) on Site-Directed deletion of four nucleotides of STAT1 binding sites.

Article Snippet: Antibodies used for Western blot were FAT1 antibody (NB100-2693; 1:500; 508 kDa), STAT1 antibody (9172S,1:1000), PDCD4 (D29C6,1:1000) (Cell Signaling technologies), β-Actin (AB8227; 1:5000; 42kDa).

Techniques: Luciferase, Activity Assay, Knockdown, Binding Assay

Journal: bioRxiv

Article Title: The atypical cadherin FAT1 is a novel regulator of STAT1, driving its pro-tumorigenic effect via the STAT1/PDCD4 axis in glioblastoma

doi: 10.1101/2023.09.22.558772

Figure Lengend Snippet: A proposed model showing a novel oncogenic signalling axis between FAT1-STAT1 in downregulating PDCD4, increasing expression of pro-tumorigenic inflammatory cytokines COX-2, IL6 and IL1B and increasing EMT, Invasion and migration.

Article Snippet: Antibodies used for Western blot were FAT1 antibody (NB100-2693; 1:500; 508 kDa), STAT1 antibody (9172S,1:1000), PDCD4 (D29C6,1:1000) (Cell Signaling technologies), β-Actin (AB8227; 1:5000; 42kDa).

Techniques: Expressing, Migration