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

Journal: Molecular Therapy. Nucleic Acids

Article Title: IL-10 modified mRNA monotherapy prolongs survival after composite facial allografting through the induction of mixed chimerism

doi: 10.1016/j.omtn.2023.02.016

Figure Lengend Snippet: IL-10 modRNA transcript produces functional protein in vitro (A–D) IL-10 modRNA produced IL-10 protein in mouse embryonic fibroblasts (MEFs), C2C12 mouse muscle myoblasts, human embryonic kidney 293 (HEK293) cells, and Jurkat cells (human T cells) at different time points by IL-10 ELISA. MEF (2 × 10 4 ), C2C12 (1 × 10 5 ), and 293 cells (1 × 10 5 ) were transfected with 100 ng modRNA. Jurkat cells (5 × 10 5 ) were transfected with 200 ng modRNA. All transfection experiments were performed in 96-well flat plate using Lipofectamine 3000 reagent. (E) Jurkat cells expressed IL-10 protein when transfected with IL-10 modRNA. Jurkat cells (12 × 10 6 ) were transfected without or with 5,000 ng IL-10 modRNA in 6-well plate. Cells were harvested one day after transfection, and 20 μg protein extract was analyzed using western blot assay. β-Actin was used as loading control. (F and G) Jurkat cells treated with IL-10 modRNA secreted functional IL-10 protein, which inhibited IL-2 and IFN-γ production in vitro . Jurkat cells (3 × 10 5 cells per well) transfected with 200 ng IL-10 modRNA were seeded in the top chambers of Transwell 96-well plates. Splenocytes (SPL; 3 × 10 5 cells per well) were seeded in bottom reservoir and stimulated with coated anti-CD3 (2 μg/mL) and soluble anti-CD28 (2 μg/mL). Supernatants in bottom reservoir were collected at different time points and analyzed using IL-2 and IFN-γ ELISA. Statistical data are represented as mean ± SD. The IL-10 modRNA group was compared with the buffer group at different time points (∗∗p < 0.005 and ∗∗∗p < 0.0005, Student’s t test).

Article Snippet: Primary antibodies were used against CD3 (SP7 clone; dilution 1:100; GeneTex) or FoxP3 (FJK-16 s clone; dilution 1:200; Thermo Fisher Scientific for 60 min.

Techniques: Functional Assay, In Vitro, Produced, Enzyme-linked Immunosorbent Assay, Transfection, Western Blot, Control

Journal: Molecular Therapy. Nucleic Acids

Article Title: IL-10 modified mRNA monotherapy prolongs survival after composite facial allografting through the induction of mixed chimerism

doi: 10.1016/j.omtn.2023.02.016

Figure Lengend Snippet: Degree of lymphocytic infiltration into facial allograft (A) Macroscopic and histological changes (indicated by hematoxylin and eosin staining) in facial OMC allografts of IL-10 modRNA and buffer groups on PODs 10–14. Facial OMC allografts from allotransplanted mice receiving single 10 μg dose of IL-10 modRNA or saline buffer on POD 3 were analyzed. Infiltrating lymphocytes into skin and muscle layers of facial OMC allografts are shown by dense violet staining. Histological Banff classification of facial OMC allografts between buffer and IL-10 modRNA groups is as shown in the right of (A). Each group contains four mice. Scale bars, 50 μm. (B) Immunohistochemistry in facial OMC allografts of IL-10 modRNA and buffer groups on PODs 10–14. Infiltrating CD3 + T cells or FoxP3 + cells into skin and muscle layers of facial OMC allografts are shown by red staining. Because CD3 is expressed in the cell surface of T cells, the staining displays a circle red. FoxP3 is a critical transcription factor and marker for mouse CD4 + CD25 + natural Tregs, and the staining displays a dense red. Scale bars, 50 μm.

Article Snippet: Primary antibodies were used against CD3 (SP7 clone; dilution 1:100; GeneTex) or FoxP3 (FJK-16 s clone; dilution 1:200; Thermo Fisher Scientific for 60 min.

Techniques: Staining, Saline, Immunohistochemistry, Marker