Journal: Nature Communications

Article Title: NR2F6 deletion revives CAR-T cell function and induces antigen-agnostic immune memory in solid tumors

doi: 10.1038/s41467-026-69796-0

Figure Lengend Snippet: A Experimental setup and timeline. Created in BioRender. Klepsch, V. ( https://BioRender.com/772xola ). Average tumor growth curve ( B ) and survival curve ( C ) in immunodeficient Rag1 −/− recipient mice. Rag1 −/− mice injected subcutaneously with 5 × 10 5 Panc02-EpCAM (100% positive) cells and treated 2 days later with 1 × 10 6 genetically modified 28ζ CAR-T cells using CRISPR/Cas9 (NTC vs Nr2f6 crispr−/− . g04 or Nr2f6 crispr−/− . g03 ). Tumor growth and survival was compared to the no therapy group that received PBS. Tumor volume was measured by caliper. n = 6 mice for Nr2f6 crispr−/−g03 and PBS and n = 5 mice for NTC and Nr2f6 crispr−/− . g04 , representative of two independent experiments. B two-way ANOVA Nr2f6 crispr−/− . g04 vs. Nr2f6 crispr−/− . g03 : ns = p = 0.9889, C overall ns = p = 0.2295, PBS vs Nr2f6 crispr−/− . g04 ns = p = 0.1213. D Experimental setup and timeline for FACS and ELISpot analysis. Ly5.1 mice were used as recipients to track Ly5.2 NTC or Nr2f6 crispr−/−.g03 CAR-T cells. Created in BioRender. Klepsch, V. ( https://BioRender.com/oeq94aw ). E Flow cytometric analysis of splenocytes from three NTC and three Nr2f6 crispr−/−.g03 CAR-T cell-treated wild type tumor-bearing mice on d8 after tumor inoculation (1 × 10 6 Panc02-EpCAM mosaic ). Increased proportion of cDC1 in Nr2f6 crispr−/−.g03 CAR T cell treated animals compared to NTC. cDC1 were gated as exemplified in Supplementary Fig. , n = 3 mice per group. F IL12p40 concentration in serum of NTC and Nr2f6 crispr−/− CAR T treated tumor-bearing mice d6 after ACT determined by LUMINEX technology. Data was normalized to NTC CAR T receiving mice. n = 2 PBS, 3 NTC CAR, and 6 Nr2f6 crispr−/− CAR mice. G , H Granzyme B ELISpot assay. Immunocompetent wild type mice were injected subcutaneously with 1 × 10 6 Panc02-EpCAM mosaic cells and left untreated (PBS) or treated 2 days later with 3.6 × 10 6 genetically modified 28ζ NTC or Nr2f6 crispr−/− . g03 CAR-T cells. Assay was performed with isolated endogenous CD3 + T cells on day 21 after tumor inoculation. NTC versus Nr2f6 crispr−/− . g03 CAR-T. n = 5 mice per group from 2 independent experiments. E – G two-tailed unpaired Student’s t-test. Data shown as mean ± SEM.

Article Snippet: For antigen presentation, mouse bone-marrow-derived dendritic cells (BMDCs) were generated as described elsewhere and either left unstimulated or stimulated with an activating antibody against CD3 (Mabtech, Nacka Strand, Sweden, 3605-1-50) as positive control as well as a set of peptides and lysates.

Techniques: Injection, Genetically Modified, CRISPR, Enzyme-linked Immunospot, Concentration Assay, Luminex, Isolation, Two Tailed Test

Journal: bioRxiv

Article Title: Multimodal Profiling of Repair-associated Immune Dynamics in a Mouse Model of Menstruation

doi: 10.64898/2026.02.06.704418

Figure Lengend Snippet: (A) Population restricted FC analysis to identify CD45 + Ly6G − NK1.1 − CD19 − CD3 + T cells in uterine tissue digests. (B) Bar plot, FC quantification: Relative abundance of T cells calculated as a percentage of total CD45 + cells per uterine horn. (C) Bar plot, FC quantification: Absolute counts of T cells per uterine horn. (D) Immunohistochemistry: detection of CD3 + T cells (black arrows) in uterine tissues 24- and 48 hr after progesterone withdrawal (representative images: 24 hr n=5, 48 hr n=6, scale bar= 50µm). (E) Population restricted FC analysis to identify CD45 + Ly6G − CD3 − CD19 − NK1.1 + NK cells in uterine tissue digests. (F) Bar plot, FC quantification: Relative abundance of NK cells calculated as a percentage of total CD45 + cells per uterine horn. (G) Bar plot, FC quantification: Absolute counts of NK cells per uterine horn. (H) Immunohistochemistry: detection of DBA-bound NK cells in uterine tissues 24- and 48 hr after progesterone withdrawal (representative images: 24 hr n=4, 48 hr n=4, scale bar= 50µm). (I) Population restricted FC analysis to identify CD45 + Ly6G − CD3 − NK1.1 − CD19 + B cells in uterine tissue digests (J) Bar plot, FC quantification: Relative abundance of B cells calculated as a percentage of total CD45 + cells per uterine horn. (K) Bar plot, FC quantification: Absolute counts of B cells per uterine horn. (L) Immunohistochemistry: detection of B220 + B cells (black arrows) in uterine tissues 24- and 48 hr after progesterone withdrawal (representative images: 24 hr n=4, 48 hr n=4, scale bar= 50µm). (FC analysis groups: control (n=15), 0 hr (prior to breakdown, n=17), 12 hr (tissue breakdown, n=22), 24 hr (tissue repair, n=16) and 48 hr (tissue remodelling, n=10) after progesterone withdrawal; statistical comparisons were made using Kruskal-Wallis tests with multiple comparisons. * p< 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001).

Article Snippet: Primary antibodies against CD3 (1:250 dilution; NB600-1441, Novus Biologicals), B220 (1:250 dilution; 14-0452-82, Invitrogen), CD14 (1:6000 dilution; ab221678, Abcam), CD64 (1:6000 dilution; 50086-R008, Sino Biological), Ly6G (1:10000 dilution; 127649, Biolegend) and DBA-lectin (1:1000 dilution; B-1035, Vector) were used in this study.

Techniques: Immunohistochemistry, Control

Journal: bioRxiv

Article Title: Multimodal Profiling of Repair-associated Immune Dynamics in a Mouse Model of Menstruation

doi: 10.64898/2026.02.06.704418

Figure Lengend Snippet: Immunohistochemistry: detection of (A) CD3+ T cells, (B) DBA-lectin+ NK cells and (C) B220+ B cells in uterine tissue sections at 0hr (prior to breakdown, n=4), 12hr (tissue breakdown, n=4-6), 24hr (tissue repair, n=4-5) and 48hr (tissue remodelling, n=4-6) hr after progesterone withdrawal (representative images, scale bar=500µm).

Article Snippet: Primary antibodies against CD3 (1:250 dilution; NB600-1441, Novus Biologicals), B220 (1:250 dilution; 14-0452-82, Invitrogen), CD14 (1:6000 dilution; ab221678, Abcam), CD64 (1:6000 dilution; 50086-R008, Sino Biological), Ly6G (1:10000 dilution; 127649, Biolegend) and DBA-lectin (1:1000 dilution; B-1035, Vector) were used in this study.

Techniques: Immunohistochemistry

Journal: bioRxiv

Article Title: Multimodal Profiling of Repair-associated Immune Dynamics in a Mouse Model of Menstruation

doi: 10.64898/2026.02.06.704418

Figure Lengend Snippet: (A) Population restricted FC analysis to identify CD45 + CD3 − CD19 − NK1.1 − CD11b + Ly6G − SiglecF − CD64 − Ly6C + MHCII − monocytes in uterine tissue digests. (B) Bar plot, FC quantification: Relative abundance of monocytes calculated as a percentage of total CD45 + cells per uterine horn. (C) Bar plot, FC quantification: Absolute counts of monocytes per uterine horn. (D) Population restricted FC analysis of CD45 + CD3 − CD19 − NK1.1 − CD11b + Ly6G − SiglecF − CD64 + Ly6C − F4/80 + MHCII + macrophages in uterine tissue digests. (E) Bar plot, FC quantification: Relative abundance of macrophages calculated as a percentage of total CD45 + cells per uterine horn. (F) Bar plot, FC quantification: Absolute counts of macrophages per uterine horn. (G) Population restricted FC analysis of CD45 + CD3 − CD19 − NK1.1 − CD11b + Ly6G + SiglecF − neutrophils in uterine tissue digests. (H) Bar plot, FC quantification: Relative abundance of neutrophils calculated as a percentage of total CD45 + cells per uterine horn. (I) Bar plot, FC quantification: Absolute counts of neutrophils per uterine horn. (FC analysis groups: control (n=15), 0 hr (prior to breakdown, n=17), 12 hr (tissue breakdown, n=22), 24 hr (tissue repair, n=16) and 48 hr (tissue remodelling, n=10) after progesterone withdrawal; statistical comparisons were made using Kruskal-Wallis tests with multiple comparisons. * p< 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001). (J-K) Immunohistochemical detection of CD14 + monocytes (cyan), CD64 + macrophages (yellow) and Ly6G + neutrophils (magenta) (merged and split channel) in uterine tissue cross sections at (J) 24 hr (repair, n=5) and (K) 48 hr (remodelling, n=3) following progesterone withdrawal (merged and split channel representative images, scale bar = 1000µm).

Article Snippet: Primary antibodies against CD3 (1:250 dilution; NB600-1441, Novus Biologicals), B220 (1:250 dilution; 14-0452-82, Invitrogen), CD14 (1:6000 dilution; ab221678, Abcam), CD64 (1:6000 dilution; 50086-R008, Sino Biological), Ly6G (1:10000 dilution; 127649, Biolegend) and DBA-lectin (1:1000 dilution; B-1035, Vector) were used in this study.

Techniques: Control, Immunohistochemical staining

Journal: bioRxiv

Article Title: Multimodal Profiling of Repair-associated Immune Dynamics in a Mouse Model of Menstruation

doi: 10.64898/2026.02.06.704418

Figure Lengend Snippet: (A) Population restricted FC analysis to identify CD45+ CD3- CD19- NK1.1- Ly6G- CD64- CD11c+ MHCII+ dendritic cells (DCs) in uterine tissue digests. (B) Bar plot, FC quantification: Relative abundance of DCs calculated as a percentage of total CD45+ cells per uterine horn. (C) Bar plot, FC quantification: Absolute counts of DCs per uterine horn. (D) Population restricted FC analysis to CD45+ CD3- CD19- NK1.1- CD11b+ Ly6G- SiglecF+ eosinophils in uterine tissue digests. (E) Bar plot, FC quantification: Relative abundance of eosinophils calculated as a percentage of total CD45+ cells per uterine horn. (F) Bar plot, FC quantification: Absolute counts of eosinophils per uterine horn. (FC analysis groups: control (n=15), 0hr (prior to breakdown, n=17), 12hr (tissue breakdown, n=22), 24hr (tissue repair, n=16) and 48hr (tissue remodelling, n=10) after progesterone withdrawal; statistical comparisons were made using Kruskal-Wallis tests with multiple comparisons. and * p< 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001).

Article Snippet: Primary antibodies against CD3 (1:250 dilution; NB600-1441, Novus Biologicals), B220 (1:250 dilution; 14-0452-82, Invitrogen), CD14 (1:6000 dilution; ab221678, Abcam), CD64 (1:6000 dilution; 50086-R008, Sino Biological), Ly6G (1:10000 dilution; 127649, Biolegend) and DBA-lectin (1:1000 dilution; B-1035, Vector) were used in this study.

Techniques: Control