Journal: bioRxiv
Article Title: LAG3 marks activated but hyporesponsive NK cells
doi: 10.1101/2024.12.18.629184
Figure Lengend Snippet: (A) C57BL/6 mice (n=4 for each day) were injected intraperitoneally with 200ug of Poly (I:C) and sacrificed in kinetic. The percentage of Sca1+, LAG3+ and CD69+ splenic NK cells was analyzed by flow cytometry. (B) The percentage of LAG3+ among Sca1+ and Sca1-NK cells on Day2 was analyzed by flow cytometry (n=5). (C-G) C57BL/6 mice (n=2 for each experiment) were injected intraperitoneally with 200ug of Poly (I:C) and sacrificed on Day2. Splenic NK cells were stained for the phospho-epitope pS6 Ser235/236 (C) and for pAkt S473 (D). Overlays of representative histograms are shown (left panels). The MFI values are indicated on the graph and represented on the right panel for each individual, n=6, 3 experiments in total. (E) NK cells were stained with Mitotracker Green and CellRox. The MFI values for each mouse (n=7, 3 experiments in total) for the analyzed marker are represented. (F) The MFI of CD98 (left) or CD71 (right) was determined by flow cytometry on LAG3+ and LAG3-NK cells (n=3). Due to the small sample size, we did not calculate p-values. (G) SSC-A and FSC-A parameters on NK cells were measured by flow cytometry in LAG3- and LAG3+ NK cells (n=5). (H) Splenic NK cells from C57BL/6 mice (n=6, 3 experiments) were isolated, stained with Cell Trace Violet (CTV) and cultured with 100ng/mL of IL-15 for 3 days. The MFI of CTV was analyzed by flow cytometry and the overlays of representative histograms are shown (left panel). The percentage of divided NK cells for each individual was calculated using the proliferation modeling tool on the FlowJo software. All the data were analyzed using a paired non-parametric Wilcoxon test. p-values are indicated on each graph.
Article Snippet: Isolated NK cells were stained with 0.6µM Invitrogen CellTrace TM Violet (CTV) Cell Proliferation Kit according to manufacturer’s instructions.
Techniques: Injection, Flow Cytometry, Staining, Marker, Isolation, Cell Culture, Software