Extended Data Fig. 3: NR increases NAD+ levels in NaLa-treated PBMC-derived NK cells.
From: Targeting lactylation reinforces NK cell cytotoxicity within the tumor microenvironment
(a) TEM showed mitochondrial morphology of activated healthy donor (HD) NK cells stimulated with NaLa (25 mM), or co-stimulated with NR (1 mM), or co-stimulated with HKL (100 μM), or co-stimulated with NR plus HKL for 24 h. The bottom-row images are magnified views of the areas in red dashed boxes above. Scale bars, 1 μm. The image is representative of three independent experiments. (b) Length of each mitochondrion (left) in each group (Each dot = 1 mitochondrion). Quantification (middle) and frequency (right) of fractured mitochondria in a single field of each group (Each dot = a single field). n = 8 donors. (c) Representative immunofluorescence images (left) and quantification (right) of mCherry-FiNad levels in naive healthy human NK cells with NaLa alone or co-stimulated with NR. Each dot represents a single field. Scale bars, 50 μm. n = 5 donors. (d) Representative immunofluorescence images (left) and quantification (right) of mCherry-FiNad levels in activated healthy human NK cells with NaLa alone or co-stimulated with NR. Each dot represents a single field. Scale bars, 50 μm. n = 5 donors. (e) Flow cytometry analysis of the percentage of Annexin V+ primary AML blasts (target cells) co-cultured for 4 h with activated healthy human NK cells subjected to various treatments (1st row); proportion of CD107a+ NK cells (2nd row), IFN-γ+ NK cells (3rd row) and Granzyme B+ NK cells (4th row) within the total population of activated healthy human NK cells with various treatments and were co-cultured with primary AML blasts. n = 8 donors. (f) Flow cytometry analysis of NKG2D, CD160 and CD38 expression on activated healthy human NK cells with indicated conditions. n = 8 donors. The data in b-f were analyzed by one-way ANOVA with Tukey’s multiple comparisons test; means ± SD.
