Journal: Nature Communications

Article Title: The signalling receptor MCAM coordinates apical-basal polarity and planar cell polarity during morphogenesis

doi: 10.1038/ncomms15279

Figure Lengend Snippet: ( a ) Upper panel, cartoon of MCAM-BD 1–3. Lower panel, yeast zygotes obtained after mating the bait strain containing pGBKT7-MCAM with the library strain containing pGADT7-FGF2, FGF4, and FGF8. BD, DNA-binding domain; AD, activation domain; +, positive control with p53-BD; and −, negative control with empty AD vector and BD-lambda. ( b ) Co-immunoprecipitation of MCAM/FGF4 and FGFR1/FGF4 with the protein lysate treated with or without the heparinase I and heparinase III (0.06 IU ml −l ). ( c ) Kinetic dissociation constant ( K D ) of FGF4/MCAM, FGF4/FGFR1, or FGF4/ FGFR2 complexes was measured using a surface plasmon resonance method. ( d ) Distribution of polarized MCAM and unpolarized FGFR1 on chemotaxing cells. The source concentration of FGF4 in the chemotaxis assay is 10 ng ml −l . ( e ) Co-localization of endogenous MCAM and the apical marker aPKCζ in chemotaxing cells. Scale bar, 20 μm. ( f , g ) Time-lapse live-cells imaging of endogenous MCAM ( f ) or exogenous MCAM-RFP ( g ) at the leading edge of chemotaxing cells.

Article Snippet: The recombinant human proteins of FGF2 (10014-HNAE), FGFR1/Fc (10616-H03H), FGFR2/Fc (10824-H03H), MCAM/Fc (10115-H02H), and IgG1 Fc (10702-HNAH) were from Sino Biological Inc.

Techniques: Binding Assay, Activation Assay, Positive Control, Negative Control, Plasmid Preparation, Immunoprecipitation, SPR Assay, Concentration Assay, Chemotaxis Assay, Marker, Imaging

Journal: Cell & Bioscience

Article Title: Oncogenic activation revealed by FGFR2 genetic alterations in intrahepatic cholangiocarcinomas

doi: 10.1186/s13578-023-01156-7

Figure Lengend Snippet: Study design. 474 ICCs were included in this study. Among 474 cases, 290 ICCs were analyzed both by FISH and NGS testing (DNA-based and RNA-based) for FGFR2 genetic alterations, and the remaining 184 cases were only analyzed by FISH. In total, there were thirty patients with FGFR2 fusion/translocation and four patients with FGFR2 genetic mutations in our cohort. Further, we selected 1534 ICC patients in 10 studies from public databases and found 38 FGFR2 genetic alterations excluding fusions in 36 patients. In addition, we also collected 31 cases with FGFR2 in-frame deletions and 65 cases with FGFR2 site mutations across multiple cancers. Finally, we depicted the comutation plot of these FGFR2 translocation/fusion, in-frame deletion and site mutation cases

Article Snippet: RBEwere infected with recombinant lentiviruses expressing different FGFR2 fusion mutants, then were distributed into 96-well plates with indicated concentrations of BGJ398 (#T1975, TargetMol, USA).

Techniques: Next-Generation Sequencing, Translocation Assay, Mutagenesis

Journal: Cell & Bioscience

Article Title: Oncogenic activation revealed by FGFR2 genetic alterations in intrahepatic cholangiocarcinomas

doi: 10.1186/s13578-023-01156-7

Figure Lengend Snippet: FGFR2 lesion spectrum in a large cohort of ICC and pan-cancer patients. In a total of 290 ICCs, five FGFR2 genetic alterations (except for translocation/fusion) were found, the types of genomic alterations were color coded and the length of the bar represents the frequency of mutations ( A ). In total of 1534 ICCs from ten studies, 38 FGFR2 genetic alterations excluding fusions were found and the types of genomic alterations were color coded ( B ). A consortium of 91,129 multiple tumors were collected to identify FGFR2 in-frame deletions. In total, eleven tumor types that carried FGFR2 genetic short in frame deletions were identified with ICC at the highest frequency (0.62%, 7/1122), notched rectangles represent the frame deletion represented twice, the percentages represent the frequency of the mutation ( C ). Pan-cancer study of 9999 cases from public database was analyzed to evaluate the prevalence of FGFR2 site mutations, there were 70 site mutations in 65 patients were found, and in ICC the prevalence was 1.08% ( D ). Other tumors that may show scattered FGFR2 point mutations but are not listed in the figure include: cancer of unknown (1/120, A511T), uterine corpus endometrial carcinoma (1/61, S252W), extrahepatic cholangiocarcinoma (1/351, R255W), gallbladder carcinoma (1/240, N441S), gastric cancer (1/866, K399Q), ovarian cancer(1/261, S252W) and urothelial carcinoma (1/96,Q259L)

Article Snippet: RBEwere infected with recombinant lentiviruses expressing different FGFR2 fusion mutants, then were distributed into 96-well plates with indicated concentrations of BGJ398 (#T1975, TargetMol, USA).

Techniques: Translocation Assay, Mutagenesis

Journal: Cell & Bioscience

Article Title: Oncogenic activation revealed by FGFR2 genetic alterations in intrahepatic cholangiocarcinomas

doi: 10.1186/s13578-023-01156-7

Figure Lengend Snippet: Histopathological features of FGFR2 genetic alteration ICCs. FGFR2 gene fusion/translocation ICCs were enriched for specific subtypes in small duct cholangiocarcinoma (CLC) with specific immunological features showing MUC5A negativity, MUC6 negativity or sporadic positivity, and CD56 positivity. HE and IHC staining are presented at 200×, the scale represents 100 µm ( A ). ICCs with FGFR2 site mutations, in-frame deletion and frame-shift deletion presented large duct (LD) or small duct (SD) in histological subtypes. HE staining are presented at 200×, the scale represents 100 µm ( B )

Article Snippet: RBEwere infected with recombinant lentiviruses expressing different FGFR2 fusion mutants, then were distributed into 96-well plates with indicated concentrations of BGJ398 (#T1975, TargetMol, USA).

Techniques: Translocation Assay, Immunohistochemistry, Staining

Journal: Cell & Bioscience

Article Title: Oncogenic activation revealed by FGFR2 genetic alterations in intrahepatic cholangiocarcinomas

doi: 10.1186/s13578-023-01156-7

Figure Lengend Snippet: Underlying co-mutation features of different types of FGFR2 genetic alterations. Comutation plot of seventeen FGFR2 translocation/fusion cases ( A ), thirty-one FGFR2 in-frame deletion cases ( B ) and sixty-five FGFR2 site mutation cases ( C )

Article Snippet: RBEwere infected with recombinant lentiviruses expressing different FGFR2 fusion mutants, then were distributed into 96-well plates with indicated concentrations of BGJ398 (#T1975, TargetMol, USA).

Techniques: Mutagenesis, Translocation Assay

Journal: Cell & Bioscience

Article Title: Oncogenic activation revealed by FGFR2 genetic alterations in intrahepatic cholangiocarcinomas

doi: 10.1186/s13578-023-01156-7

Figure Lengend Snippet: Various oncogenic activities and diverse responses to FGFR-selective small molecule kinase inhibitor (SMKI) among FGFR2 mutants. Proliferation activities of AML12 cells expressing MCS, FGFR2 and different FGFR2 mutants are shown ( A ). Representative images of transwell migration assay and average numbers of migrated AML12 cells expressing FGFR2 and different FGFR2 mutants are shown ( B ), the scale represents 100 µm. Representative images of invasion assay and average number of invasive AML12 cells expressing FGFR2 and various FGFR2 mutants are shown ( C ). Cellular skeleton staining revealed the morphological changes in AML12 cells when expressing Lenti-CMV-MCS control virus (MCS), FGFR2 and different FGFR2 mutants ( D ). Evaluation the sensitivity of RBE cells with different FGFR2 mutants to BGJ398 ( E )

Article Snippet: RBEwere infected with recombinant lentiviruses expressing different FGFR2 fusion mutants, then were distributed into 96-well plates with indicated concentrations of BGJ398 (#T1975, TargetMol, USA).

Techniques: Expressing, Transwell Migration Assay, Invasion Assay, Staining, Control, Virus

Journal: Cell & Bioscience

Article Title: Oncogenic activation revealed by FGFR2 genetic alterations in intrahepatic cholangiocarcinomas

doi: 10.1186/s13578-023-01156-7

Figure Lengend Snippet: Frameshift mutation resulting in premature stop codons is also a potential therapeutic target. Proliferation activities of AML12 cells expression Lenti-CMV-MCS control virus (MCS), FGFR2, FGFR2-BICC1 fusion and FGFR2 I548Wfs*8 mutant are shown ( A ). Representative images of transwell migration and average numbers of migrated cells expressing above three clones ( B ), the scale represents 100 µm. Representative images of invasion and average colonies of invasion cells expressing above three clones in AML12 cells are shown ( C ). Capability of FGFR2 I548Wfs*8 rendering RBE cells sensitive to BGJ398 ( D )

Article Snippet: RBEwere infected with recombinant lentiviruses expressing different FGFR2 fusion mutants, then were distributed into 96-well plates with indicated concentrations of BGJ398 (#T1975, TargetMol, USA).

Techniques: Mutagenesis, Expressing, Control, Virus, Migration, Clone Assay