Journal: mAbs

Article Title: Inhibition of HER3 activation and tumor growth with a human antibody binding to a conserved epitope formed by domain III and IV

doi: 10.1080/19420862.2017.1319023

Figure Lengend Snippet: Expression levels (receptors per cell) of EGFR, HER2, and HER3 on various cancer cell lines and binding of IgG-43 as analyzed by flow cytometry.

Article Snippet: A549 cells were obtained from CLS Cell Lines Services (Eppelheim, Germany).

Techniques: Expressing, Binding Assay, Flow Cytometry

Journal: mAbs

Article Title: Inhibition of HER3 activation and tumor growth with a human antibody binding to a conserved epitope formed by domain III and IV

doi: 10.1080/19420862.2017.1319023

Figure Lengend Snippet: Inhibition of ligand-dependent and ligand-independent proliferation. (a) Inhibition of proliferation of MCF-7, NCI-N87, and A549 cells incubated for one week under low (0.2%) serum concentrations in the presence of 10 ng/ml heregulin and either IgG 3–43 (3–43) or rituximab (Control) as a negative control antibody. (b) Inhibition of proliferation of FaDu cells without the addition of heregulin under the same conditions as in (a). (c) Colony formation assay. SKBR3 or BT474 were grown in 12-well plates (1,000 cells per well) and incubated with IgG 3–43 (50 nM) or trastuzumab (Tras.) for 12 d. Medium and antibodies were exchanged after 7 d. Cells incubated without antibody were included as a control (con). Shown are crystal violet-stained wells and the quantification of 2 independent experiments performed with triplicate wells.

Article Snippet: A549 cells were obtained from CLS Cell Lines Services (Eppelheim, Germany).

Techniques: Inhibition, Incubation, Negative Control, Colony Assay, Staining

Journal: Nature Methods

Article Title: Optimal precision and accuracy in 4Pi-STORM using dynamic spline PSF models

doi: 10.1038/s41592-022-01465-8

Figure Lengend Snippet: Mic60 is a component of the MICOS complex, and is involved in the formation and maintenance of crista junctions that connect the crista membrane with the inner boundary membrane. a , Mic60 in a U-2 OS cell, labeled with primary and secondary antibodies. The Mic60 signals appear as structured, punctate clusters. The localizations are color coded according to their z coordinate (identical color scales in a – d ). Scale bar, 200 nm. b , Magnified view of the boxed region in a . Scale bar, 50 nm. c , Mic60 in a COS-7 cell, in which the crista junctions exhibit a linear organization over segments of the inner boundary membrane. Scale bar, 200 nm. d , Magnified view of the boxed region in c . Scale bar, 50 nm. e , f , Unwrapped views of the Mic60 localization density around the surface of the mitochondria, showing the nanoscale distribution of Mic60. In U-2 OS cells, Mic60 appears predominantly punctate, with pairs or clusters of signal density separated by 20–40 nm (Extended Data Fig. and Supplementary Fig. ). In COS-7 cells, Mic60 appears to have a zigzag or double-line arrangement, with a typical width of approximately 25 nm (Extended Data Fig. and Supplementary Fig. ). Dashed lines indicate the extent of the data in f . g , Two-color image of Mic60 (blue) and mitochondrial nucleoids (yellow) in a COS-7 cell, stained with antibodies labeled with Alexa Fluor 647 and Cy5.5, respectively. Scale bar, 1 µm. h , Detailed view of the boxed region in g . Lower density of Mic60 close to the DNA signal, suggesting fewer crista junctions in these regions. i , Cross-section ( x – z ) through the region indicated by the dashed lines in h , showing Mic60 at the inner boundary membrane, and a DNA cluster in the center of the mitochondrion. j , A 3D perspective view of the mitochondrion shown in h and i , where the Mic60 and DNA signals have been rendered as isosurfaces. Scale bars, 250 nm ( h – j ).

Article Snippet: Experiments were performed using either standard COS-7 cells or U-2 OS cells obtained from American Type Culture Collection (ATCC), or gene-edited U-2 OS cells expressing a SNAP-tagged version of the nucleoporin Nup107 (CLS Cell Lines Service, U-2OS-ZFN-SNAP-Nup107 clone 294) or Nup96 (CLS Cell Lines Service, U-2OS-CRISPR-NUP96-SNAP clone 33) .

Techniques: Labeling, Staining

Journal: Nature Communications

Article Title: A topological refactoring design strategy yields highly stable granulopoietic proteins

doi: 10.1038/s41467-022-30157-2

Figure Lengend Snippet: A Dose–response curves of NFS-60 proliferation upon 48 h treatment with Filgrastim (rhG-GCSF), Boskar3, or Boskar4. Datapoints and error bars represent mean ± standard deviations of three biologically independent replicates. B Time-dependent proliferation trajectory of surface-immobilised NFS-60 cells over a 10-day treatment. Data points and shades represent mean and standard deviation values of three biologically independent replicates. Experiments were performed three times in triplicates. Data of one representative experiment is shown. C , D Dose- and time-dependent proliferation trajectories over a 5-day treatment of free-floating NFS-60 cells, under the influence of Boskar3 and Boskar4 treatments, respectively. Data points and shades represent mean and standard deviation values of three biologically independent replicates. E G-CSFR-deficient primary stem cells (G-CSFR KO), show abolished proliferative responses to either rhG-CSF or the designs. Experiment was performed twice in triplicates. Data points and shades represent mean and standard deviation values of three biologically independent replicates. F Intracellular levels of phospho-AKT (Thr308), phospho-ERK1/2 (p44/42 MAPK), phospho-STAT3 (Tyr705), and phospho-STAT5 (Tyr694) in CD34 + HSPCs treated with rhG-CSF or the designs (see the “Methods” section). Geometric mean of the expression intensity of each phospho-protein (GeoMean intensity) is shown on the y -axis. The experiment was performed twice.

Article Snippet: NFS-60 cells were cultured in IL-3-containing RPMI 1640 medium, supplemented with L-glutamine, 10% KMG-5 and 10% FBS (CLS, cell line services).

Techniques: Standard Deviation, Expressing

Journal: Nature Communications

Article Title: A topological refactoring design strategy yields highly stable granulopoietic proteins

doi: 10.1038/s41467-022-30157-2

Figure Lengend Snippet: The creation of two-copy Boskar4 tandems with different flexible spacers, through A a 24-residue-linker (Boskar4_t2), and B a short 6-residue-linker (Boskar4_st2). C Dose–response curves in NFS-60 cells show Boskar4_t2 (EC 50 = 180 pM), and Boskar4_st2 (EC 50 = 7.6 pM) to be 11- and 263-fold more active than the single domain Boskar4, respectively. Datapoints and error bars represent mean ± standard deviations of 3 biologically independent replicates. D , E This greatly enhanced potency is also evident in the dose- and time-dependent NFS-60 proliferation kinetics, highlighting the requirement for an optimal G-CSFR dimerisation spacing to achieve maximal G-CSFR activation. The experiments in C , E were performed as described in figure legend to Fig. .

Article Snippet: NFS-60 cells were cultured in IL-3-containing RPMI 1640 medium, supplemented with L-glutamine, 10% KMG-5 and 10% FBS (CLS, cell line services).

Techniques: Activation Assay

Journal: British Journal of Cancer

Article Title: Combined proteome and transcriptome analyses for the discovery of urinary biomarkers for urothelial carcinoma

doi: 10.1038/bjc.2013.157

Figure Lengend Snippet: Classification of the proteins identified in the conditioned medium of UCB cell lines. The proteins identified in the conditioned media and in the corresponding cells were divided into categories using the GO Cellular Components function in STRING 9.0 ( www.string-db.org ). The number of proteins in each analysis was: 5637=622/546, EJ28=649/717, HB-CLS-2=822/462 (cell pellet/secretome).

Article Snippet: 5637 and HB-CLS-2 cells were purchased from CLS Cell Lines Service GmbH (Eppelheim, Germany) and EJ28 cells were a kind gift from Elizabeth Hodgkins, University of Birmingham.

Techniques: