Journal: Nature communications

Article Title: Distinctive phenotypes and functions of innate lymphoid cells in human decidua during early pregnancy.

doi: 10.1038/s41467-019-14123-z

Figure Lengend Snippet: Fig. 6 dNK responses triggered by activating KIR. a Representative XCL1 and CD107a staining by CyTOF in lineage negative (Lin−) CD56+ decidual cells co-expressing 1–3 Killer-cell immunoglobulin-like receptors (KIR) following a 6 h co-culture with K562 (n = 10). b Representative XCL1 and CD107a staining by mass cytometry in Lin-CD56+ cells co-expressing 1–3 KIR following 4 h stimulation by PMA plus ionomycin (n = 8). c Representative XCL1 staining by flow cytometry in Lin-CD56+KIR2DS4+ decidual cells co-expressing 1–3 additional KIRs following activation via P815 cells coated with anti-KIR2DS4 (n = 6). d Correlation of frequency of XCL1+Lin-CD56+ decidual cells and mean side scatter for the same subset (n = 20). Two-tailed p-value calculated for Pearson correlation coefficients. Two-tailed one-way ANOVA of matched data points with Tukey correction and 95% confidence level was used. *p < 0.05, **p < 0.01, ***p < 0.001. Source data are provided as a Source Data file.

Article Snippet: The murine cell line P815 and human cell line K562 were purchased from DSMZ in Germany.

Techniques: Staining, Expressing, Co-Culture Assay, Mass Cytometry, Cytometry, Activation Assay, Two Tailed Test

Journal: Journal of cell science

Article Title: A role for membrane-bound CD147 in NOD2-mediated recognition of bacterial cytoinvasion.

doi: 10.1242/jcs.016980

Figure Lengend Snippet: Fig. 1. The CARD domains of NOD2 interact with the intracellular stalk of CD147. (A) HEK293 cells stably transfected with NOD2 expression construct (HEKNOD2) or empty vector (HEKmock) were used for co-immunoprecipitation of CD147 and NOD2. CD147 immunocomplexes and whole cell lysates were assayed for endogenous and precipitated NOD2 and CD147. Precipitates from irrelevant antibody were used as specificity control (ctrl). (B) HEK293 cells were transfected with constructs containing Xpress-tagged NOD2 or NOD2 domains. After precipitation of endogenous CD147, co-precipitated NOD2 or NOD2 domains were visualized using anti-Xpress antibody. Only full-length NOD2 and a fusion protein containing both CARD domains were found in CD147 immmunocomplexes (black arrowheads). (C) HEK293 cells were transfected with constructs containing Xpress-tagged NOD2 or NOD2 domains in combination with a construct containing the intracellular domain of CD147 (IC) fused to EGFP. After precipitation of Xpress-tagged NOD2 domains, EGFP-CD147-IC was identified in the immunocomplexes using anti- GFP antibody. Only full-length NOD2 and both CARD domains were able to precipitate with GFP-CD147-IC (indicated by black arrowheads). Antibody alone (Ab) was used as control. (D) CD147 immunocomplexes derived from THP1 monocytes were used to verify the interaction of CD147 with endogenous NOD2. Detection of two specific bands by use of NOD2 antibody points to the presence of more than one NOD2 isoform.

Article Snippet: Human epithelial HEK293 cells (ACC305), cervical carcinoma HeLaS3 cells (ACC161), colonic carcinoma SW480 (ACC313) and human acute monocytic cell line THP1 (ACC16) were purchased from the German Collection of Microorganisms and Cell Cultures (DSMZ, Braunschweig, Germany).

Techniques: Stable Transfection, Transfection, Expressing, Construct, Plasmid Preparation, Immunoprecipitation, Control, Derivative Assay

Journal: Journal of cell science

Article Title: A role for membrane-bound CD147 in NOD2-mediated recognition of bacterial cytoinvasion.

doi: 10.1242/jcs.016980

Figure Lengend Snippet: Fig. 2. Expression and regulation of CD147. (A) CD147 mRNA levels in monocytic THP1 cells are upregulated by stimulation with MDP, TNF- plus IFN- and infection with L. monocytogenes. (B) THP1 monocytes were stimulated for 24 hours with different agents (thick line) or left untreated (thin line). Cell surface expression of CD147 protein was detected by FACS. Irrelevant antibody was used as specificity control (gray curve). A variety of proinflammatory stimuli (MDP, LPS, TNF-/IFN-) lead to an increased expression of CD147. Note that TNF- alone showed only a minor effect.

Article Snippet: Human epithelial HEK293 cells (ACC305), cervical carcinoma HeLaS3 cells (ACC161), colonic carcinoma SW480 (ACC313) and human acute monocytic cell line THP1 (ACC16) were purchased from the German Collection of Microorganisms and Cell Cultures (DSMZ, Braunschweig, Germany).

Techniques: Expressing, Infection, Control

Journal: Journal of cell science

Article Title: A role for membrane-bound CD147 in NOD2-mediated recognition of bacterial cytoinvasion.

doi: 10.1242/jcs.016980

Figure Lengend Snippet: Fig. 3. Co-localization of CD147 and NOD2 at the membrane of epithelial and myelomonocytic cells. (A) HeLaS3 cells were transiently transfected with GFP- NOD2 (green) and stained for CD147 (red) and DAPI as a DNA counterstain (blue). (B) Transfected cells were infected with L. monocytogenes (MOI=100) and fixed after 1 hour. In the basal as well as in the infected state, NOD2 and CD147 colocalize in the membrane compartment. An accumulation of both NOD2 and CD147 in areas of early bacterial invasion is detectable (open arrowheads and inset in B). (C) Indirect immunofluorescence micrographs of PMA- differentiated THP1 myelomonocytic cells. Staining with specific antibodies against endogenous NOD2 (red channel) and CD147 (green channel) shows a partial colocalization at the cell membrane (arrowheads in merged image). Images are representative of at least three independent experiments. Original magnification: 100.

Article Snippet: Human epithelial HEK293 cells (ACC305), cervical carcinoma HeLaS3 cells (ACC161), colonic carcinoma SW480 (ACC313) and human acute monocytic cell line THP1 (ACC16) were purchased from the German Collection of Microorganisms and Cell Cultures (DSMZ, Braunschweig, Germany).

Techniques: Membrane, Transfection, Staining, Infection, Immunofluorescence