Fig 2: CREM is expressed in CAR70/IL-15 NK cells upon encounter with CD70+ tumor cells and is associated with an activated phenotype.(a) CREM expression in non-transduced (NT), IL-15, CAR70/IL-15, and CAR70 NK cells as assessed by qPCR (n = 8 donors); (b) Percentage of CREM positive cells in live CD45+ GFP- CD56+ NK cells from NT, IL-15, CAR70/IL-15, and CAR70 NK cells following 24 h coculture with UMRC3 tumor cells at an effector-to-target (E:T) ratio of 1:1 as assessed by mass cytometry (n = 3 donors); (c) Violin plots of CREM expression in the various NK cell conditions at the single cell level as assessed by mass cytometry (n = 3 donors); the gray dashed line represents the median expression in NT NK cells; (d) Violin plots of CREM expression at the single cell level compared between the various clusters of the merged tSNE_CUDA analysis of the various conditions (n = 3 donors); (e) Percentage of CREM positive NK cells in the tSNE_CUDA clusters (n = 3 donors); (f) Frequencies of NK cells expressing CREM in the tSNE_CUDA clusters (colored portion); the degree of the color reflects the robust z-score of CREM expression in the corresponding cluster on a scale ranging from dark orange (z = +1) to dark blue (z = −1); (g) Total cells (blue), cells expressing the CAR (based on CD27 staining; red) and CREM (green) are shown on the tSNE_CUDA plots. Inset numbers indicate the percentages (%) of CAR expression on and CREM expression in the corresponding NK cell conditions; (h) Violin plots of expression of select markers of activation, inhibition as well as checkpoints in NK cells from clusters 1 (most abundant in CAR70/IL-15 NK cells) and 5 (most abundant in NT NK cells) at the single cell level as assessed by mass cytometry (n = 3 donors); (i) Heatmap showing expression of key NK cell phenotypic markers on fractions of positive CREM expression (CREM+) and negative CREM expression (CREM-) in the various NK cell conditions. Expression of each marker is represented by the robust z-score for the expression normalized across the conditions with a color scale ranging from blue (z = −1) to orange (z = +1) and the size of the circle for percentage expression. GZMB: granzyme B. ns: non-significant. Statistical comparisons were performed using one-way ANOVA (Fisher’s LSD test, a-e), t-tests (h), and one minus pearson hierarchical clustering (i). Data are represented as mean ± SEM. Source Data

Fig 3: Intravesical instillation of PFD@PDA NPs following SCI preserves bladder function and alleviates inflammatory cell infiltration in the bladder. a) Schematic illustration of the establishment of the 14-day SCI model in rats, the design of treatment regimens and subsequent urodynamic assessment. b) Standard urodynamic tracings and representative urodynamic tracings in each group. Standard urodynamic tracings illustrate bladder filling, baseline pressure during the storage phase, peak detrusor pressure during voiding, and active bladder contractions that lead to urine expulsion. c-h) Quantitative analysis of bladder baseline pressure (c), intermicturition interval (d), residual volume (e), infused volume (f), capacity (g) and voiding efficiency (h) in rats after different treatments. Sham group: n = 3; all other groups: n ≥ 5. i) Schematic illustration of the establishment of the 4-day SCI model in mice and the design of treatment regimens. j) Images of bladders in mice after different treatments (n = 6). k) Quantitative analysis of the bladder-to-body weight ratio across different experimental groups (n = 6). l) Quantitative analysis of bladder-infiltrating myeloid cells (CD45+CD11b+) across different treatment groups (n ≥ 4). m, p) Representative flow cytometry plots and quantitative analysis of bladder-infiltrating monocytes (CD11b+Ly6C+) across different treatment groups (n ≥ 4). n, q) Representative flow cytometry plots and quantitative analysis of bladder-infiltrating macrophages (CD11b+F4/80+) across different treatment groups (n ≥ 4). o, s) Representative flow cytometry plots and quantitative analysis of bladder-infiltrating neutrophils (CD11b+Ly6G+) across different treatment groups (n ≥ 4). r) Quantitative analysis of bladder-infiltrating M1-type macrophages (CD11b+F4/80+iNOS+) across different treatment groups (n ≥ 4). Data are presented as mean ± SD. Statistical significance was determined by using one-way ANOVA with Tukey’s multiple comparisons. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001, ns: not significant