Journal: iScience

Article Title: Altered immunometabolic response to fasting in humans living with obesity

doi: 10.1016/j.isci.2025.112872

Figure Lengend Snippet: Cell counts and T cell subsets at baseline and after a 48-h fast in individuals with L-BMI or O-BMI (A) Total white blood cell count. (B) Absolute lymphocyte counts in whole blood. (C) Proportion of white bloods cells that were lymphocytes in whole blood. (D) Proportion of isolated T cells derived from whole blood that were CD4 + /CD8 − , CD4 − /CD8 + , or double-negative (CD4 − /CD8 − ). (E) Proportion of CD4 + T cells that were either Th1, Th2, Th17, Th22, or Treg cells. Large circles or triangles are estimated marginal means and black vertical bars are 95% confidence intervals derived from a linear mixed effects model. Orange circles represent the lean BMI (L-BMI) group and teal triangles represent the obese BMI (O-BMI) group. Small circles or triangles represent individual participant data. N = 32; 8 males/8 females per BMI group. ∗Group × time interaction, p < 0.05; ‡main effect of time, p < 0.05; †main effect of group, p < 0.05; c L-BMI vs. O-BMI at baseline, p < 0.05.

Article Snippet: Treg, CD4 + , and CD8 + cells were quantified at Baseline and 48 h. First, 1 × 10 6 freshly isolated T cells were stained with CD45 VioBlue (Miltenyi Biotec, Cat. No. 130-110-637), CD4 VioGreen (Miltenyi Biotec, Cat. No. 130-113-230), CD25 VioBright B515 (Miltenyi Biotec, Cat. No. 130-113-287), CD127 PE (Miltenyi Biotec, Cat. No. 130-133-753), and CD8 PerCP-Vio700 (Miltenyi Biotec, Cat. No. 130-110-682) for 10 min in the dark at 4°C.

Techniques: Cell Counting, Isolation, Derivative Assay

Journal: iScience

Article Title: Altered immunometabolic response to fasting in humans living with obesity

doi: 10.1016/j.isci.2025.112872

Figure Lengend Snippet: Markers of T cell function before and after a 48-h fast in individuals with L-BMI or O-BMI (A) Expression of CD4 on isolated human T cells. (B) Expression of CD8 on isolated human T cells. (C) Intracellular secretion of IFNγ in isolated human T cells following a 24-h activation culture. (D) Intracellular secretion of IL-17 in isolated human T cells following a 24-h activation culture. Large circles or triangles are estimated marginal means and black vertical bars are 95% confidence intervals derived from a linear mixed effects model. Orange circles represent the lean BMI (L-BMI) group and teal triangles represent the obese BMI (O-BMI) group. Small circles or triangles represent individual participant data. N = 32; 8 males/8 females per BMI group. ∗Group × time interaction, p < 0.05; ‡main effect of time, p < 0.05; †main effect of group, p < 0.05; c L-BMI vs. O-BMI at baseline, p < 0.05. MFI, median fluorescence intensity.

Article Snippet: Treg, CD4 + , and CD8 + cells were quantified at Baseline and 48 h. First, 1 × 10 6 freshly isolated T cells were stained with CD45 VioBlue (Miltenyi Biotec, Cat. No. 130-110-637), CD4 VioGreen (Miltenyi Biotec, Cat. No. 130-113-230), CD25 VioBright B515 (Miltenyi Biotec, Cat. No. 130-113-287), CD127 PE (Miltenyi Biotec, Cat. No. 130-133-753), and CD8 PerCP-Vio700 (Miltenyi Biotec, Cat. No. 130-110-682) for 10 min in the dark at 4°C.

Techniques: Cell Function Assay, Expressing, Isolation, Activation Assay, Derivative Assay, Fluorescence

Journal: Nature Communications

Article Title: Co-delivery of sorafenib and an FSP1 inhibitor triggers dual ferroptosis in tumor cells and immunosuppressive macrophages for enhanced immunotherapy in mouse models of hepatocellular carcinoma

doi: 10.1038/s41467-025-65056-9

Figure Lengend Snippet: a Schematic illustration of the research procedure in subcutaneous HCC mouse model. b Relative tumor volume curves of different treatment groups ( n = 5 independent mice). c Animal survival after different treatments ( n = 5 independent mice). d Quantification of fluorescence intensity in Supplementary Fig. f ( n = 5 independent mice). e Immunofluorescence images of tumor tissue slices collected from different groups stained with anti-F4/80/CD206, anti-F4/80/CD86, and anti-CD11c/MHC II after different treatments ( n = 5 independent mice; Scale bar = 100 μm). f Quantification of the percentage of F4/80 + CD206 + M2 TAMs, F4/80 + CD86 + M1 TAMs, and CD11c + MHC II + actDCs in ( e ) ( n = 5 independent mice). g Schematic illustration of the research procedure in orthotopic HCC mouse model. h Relative bioluminescence intensity curves for the various treatment groups ( n = 5 independent mice). i Animal survival after different treatments ( n = 5 independent mice). j Flow cytometric analysis of anti-F4/80/CD206-stained macrophages, anti-F4/80/CD86-stained macrophages, anti-CD11c/MHC II-stained DCs, anti-CD3/CD4-stained T cells, anti-CD3/CD8-stained T cells, and anti-CD4/Foxp3-stained T cells in tumor tissues after different treatments ( n = 5 independent mice). Unless specified otherwise, error bars represent the mean ± SEM. Statistical significance was determined by one-way ANOVA with Tukey’s test ( b , d , f , h ) or log-rank test ( c , i ) and P -values were indicated. Source data are provided as a Source Data file. The elements in Fig. 7a, g were created by Adobe Illustrator.

Article Snippet: PerCP/Cyanine5.5 anti-mouse F4/80 (1:200, #E-AB-F0995J), APC anti-mouse CD206 (1:200, #E-AB-F1135E), PE/Cyanine7 anti-mouse CD86 (1:200, #E-AB-F0994H), Fluor Red 780 anti-mouse CD80 (1:200, #E-AB-F0992S), APC anti-mouse CD11c (1:200, #E-AB-F0991E), Fluor Violet 450 anti-mouse CD3 (1:200, #E-AB-F1013Q), Fluor Red 780 anti-mouse CD4 (1:200, #E-AB-F1097S), PerCP/Cyanine5.5 anti-mouse CD8 (1:200, #E-AB-F1104J), FITC anti-human/mouse CD44 (1:200, #E-AB-F1100C), PE anti-mouse Foxp3 (1:200, #E-AB-F1238D), FITC anti-mouse MHC II (1:200, #E-AB-F0990C), APC anti-mouse CD62L (1:200, #E-AB-F1011E), APC anti-mouse PD-L1 (1:200, #E-AB-F1132E), PerCP anti-human CD45 (1:200, #E-AB-F1137F), Fluor647 anti-human CD68 (1:200, #E-AB-F1299M), FITC anti-human CD206 (1:200, #E-AB-F1161C), PE anti-human CD80 (1:200, #E-AB-F1232D), APC anti-human HLA-DR (1:200, #E-AB-F1111E), PE anti-human CD11c (1:200, #E-AB-F1118D), APC anti-human CD3 (1:200, #E-AB-F1001E), and FITC anti-human CD8 (1:200, #E-AB-F1110C) were purchased from Elabscience (Wuhan, China).

Techniques: Fluorescence, Immunofluorescence, Staining

Journal: Nature Communications

Article Title: Co-delivery of sorafenib and an FSP1 inhibitor triggers dual ferroptosis in tumor cells and immunosuppressive macrophages for enhanced immunotherapy in mouse models of hepatocellular carcinoma

doi: 10.1038/s41467-025-65056-9

Figure Lengend Snippet: a Flow cytometric analysis of anti-PD-L1-stained cells in tumor tissues after different treatments ( n = 5 independent mice). b Quantification of the percentage of PD-L1 positive cells in ( a ) ( n = 5 independent mice). c Schematic diagram of the mechanism of Sv@PM-M2p-mediated upregulation of PD-L1 within HCC TME. d Schematic illustration of the research procedure in bilateral subcutaneous HCC mouse model. e Representative bioluminescence images of bilateral subcutaneous HCC mice at specific time points with different treatments ( n = 5 independent mice). f Relative bioluminescence intensity curves for the various treatment groups ( n = 5 independent mice). g Flow cytometric analysis of anti-F4/80/CD206-stained macrophages and anti-F4/80/CD86-stained macrophages in tumor tissues after different treatments ( n = 5 independent mice). h Quantification of the percentage of F4/80 + CD206 + M2 TAMs and F4/80 + CD86 + M1 TAMs in ( g ) ( n = 5 independent mice). i Flow cytometric analysis of anti-CD44/CD62L-stained T cells (gated on CD3 + CD8 + T cells) in the spleens after different treatments ( n = 5 independent mice). j Quantification of the percentage of CD44 + CD62L - memory T cells in ( i ) ( n = 5 independent mice). k The assay of IFN-γ, TNF, IL-6, and IL-12 in serum after different treatments ( n = 5 independent mice). Unless specified otherwise, error bars represent the mean ± SEM. Statistical significance was determined by one-way ANOVA with Tukey’s test ( b , f , h , j , k ) and P -values were indicated. Source data are provided as a Source Data file. The elements in Fig. 8c, d were created by Adobe Illustrator.

Article Snippet: PerCP/Cyanine5.5 anti-mouse F4/80 (1:200, #E-AB-F0995J), APC anti-mouse CD206 (1:200, #E-AB-F1135E), PE/Cyanine7 anti-mouse CD86 (1:200, #E-AB-F0994H), Fluor Red 780 anti-mouse CD80 (1:200, #E-AB-F0992S), APC anti-mouse CD11c (1:200, #E-AB-F0991E), Fluor Violet 450 anti-mouse CD3 (1:200, #E-AB-F1013Q), Fluor Red 780 anti-mouse CD4 (1:200, #E-AB-F1097S), PerCP/Cyanine5.5 anti-mouse CD8 (1:200, #E-AB-F1104J), FITC anti-human/mouse CD44 (1:200, #E-AB-F1100C), PE anti-mouse Foxp3 (1:200, #E-AB-F1238D), FITC anti-mouse MHC II (1:200, #E-AB-F0990C), APC anti-mouse CD62L (1:200, #E-AB-F1011E), APC anti-mouse PD-L1 (1:200, #E-AB-F1132E), PerCP anti-human CD45 (1:200, #E-AB-F1137F), Fluor647 anti-human CD68 (1:200, #E-AB-F1299M), FITC anti-human CD206 (1:200, #E-AB-F1161C), PE anti-human CD80 (1:200, #E-AB-F1232D), APC anti-human HLA-DR (1:200, #E-AB-F1111E), PE anti-human CD11c (1:200, #E-AB-F1118D), APC anti-human CD3 (1:200, #E-AB-F1001E), and FITC anti-human CD8 (1:200, #E-AB-F1110C) were purchased from Elabscience (Wuhan, China).

Techniques: Staining

Journal: Nature Communications

Article Title: Co-delivery of sorafenib and an FSP1 inhibitor triggers dual ferroptosis in tumor cells and immunosuppressive macrophages for enhanced immunotherapy in mouse models of hepatocellular carcinoma

doi: 10.1038/s41467-025-65056-9

Figure Lengend Snippet: a Schematic illustration of the research procedure in PDX subcutaneous mouse model. b Relative tumor volume curves of different treatment groups ( n = 5 independent mice). c Collected tumor tissues of mice at the end of treatment in different groups. d Tumor weight at the end of treatment in different groups ( n = 5 independent mice). e Immunofluorescence images of tumor tissue slices collected from different groups stained with Liperfluo after different treatments ( n = 5 independent mice; Scale bar = 100 μm). f Quantification of fluorescence intensity in ( e ) ( n = 5 independent mice). g Flow cytometric analysis of anti-CD68/CD206-stained macrophages (gated on CD45 + cells), anti-CD68/CD80-stained macrophages (gated on CD45 + cells), anti-HLA-DR/CD11c-stained DCs (gated on CD45 + Lin-1 – T cells), and anti-CD3/CD8-stained T cells (gated on CD45 + cells) in tumor tissues after different treatments ( n = 5 independent mice). h Quantification of the percentage of CD68 + CD206 + M2 TAMs, CD68 + CD80 + M1 TAMs, HLA-DR + CD11c + actDCs, and CD3 + CD8 + Tc cells in ( g ) ( n = 5 independent mice). Unless specified otherwise, error bars represent the mean ± SEM. Statistical significance was determined by one-way ANOVA with Tukey’s test ( b , d , f , h ) and P -values were indicated. Source data are provided as a Source Data file. The elements in Fig. 9a were created by Adobe Illustrator.

Article Snippet: PerCP/Cyanine5.5 anti-mouse F4/80 (1:200, #E-AB-F0995J), APC anti-mouse CD206 (1:200, #E-AB-F1135E), PE/Cyanine7 anti-mouse CD86 (1:200, #E-AB-F0994H), Fluor Red 780 anti-mouse CD80 (1:200, #E-AB-F0992S), APC anti-mouse CD11c (1:200, #E-AB-F0991E), Fluor Violet 450 anti-mouse CD3 (1:200, #E-AB-F1013Q), Fluor Red 780 anti-mouse CD4 (1:200, #E-AB-F1097S), PerCP/Cyanine5.5 anti-mouse CD8 (1:200, #E-AB-F1104J), FITC anti-human/mouse CD44 (1:200, #E-AB-F1100C), PE anti-mouse Foxp3 (1:200, #E-AB-F1238D), FITC anti-mouse MHC II (1:200, #E-AB-F0990C), APC anti-mouse CD62L (1:200, #E-AB-F1011E), APC anti-mouse PD-L1 (1:200, #E-AB-F1132E), PerCP anti-human CD45 (1:200, #E-AB-F1137F), Fluor647 anti-human CD68 (1:200, #E-AB-F1299M), FITC anti-human CD206 (1:200, #E-AB-F1161C), PE anti-human CD80 (1:200, #E-AB-F1232D), APC anti-human HLA-DR (1:200, #E-AB-F1111E), PE anti-human CD11c (1:200, #E-AB-F1118D), APC anti-human CD3 (1:200, #E-AB-F1001E), and FITC anti-human CD8 (1:200, #E-AB-F1110C) were purchased from Elabscience (Wuhan, China).

Techniques: Immunofluorescence, Staining, Fluorescence

Journal: Frontiers in Immunology

Article Title: DNase I alleviates renal inflammatory injury in MRL/lpr mice by inhibiting NETs formation

doi: 10.3389/fimmu.2025.1656069

Figure Lengend Snippet: DNase I treatment decreased the number of immune inflammatory cells in the kidneys and spleens of mice in the MRL/lpr model. (A, B) The TIMER algorithm was used to assess the proportion of immune cell infiltration in the kidneys of control (Ctr), MRL/lpr (Lpr), and DNase I-treated MRL/lpr (DNase) mice at 21 weeks of age. The percentages of CD3+CD4+ T cell subsets (C, G) , CD3+CD8+ T cell subsets (D, H) , and CD3-CD19+B cell subsets (E, I) in the spleen were quantified using flow cytometry across the normal control, MRL/lpr, and DNase I treatment MRL/lpr group. (F, J) Flow cytometric and quantitative analysis of CD4+CD25+Foxp3+ regulatory T cell (Treg) percentages in the spleens of mice from each experimental group were also conducted. Data were expressed as means ± SD for groups of three mice. * P < 0.05, ** P < 0.01, and *** P < 0.001 vs. normal control ( t test). # P < 0.05 vs. MRL/lpr mice (Bonferroni correction; two comparisons were made).

Article Snippet: The following antibodies were utilized for staining: anti-CD3 antibody (E-AB-F1013Q), anti-CD4 antibody (E-AB-F1097S), anti-CD8 antibody (E-AB-F1104J), anti-CD19 antibody (E-AB-F0986E), anti-CD25 antibody (E-AB-F1102D), and anti-Foxp3 antibody (E-AB-F1238E), all sourced from Elabscience (China).

Techniques: Control, Flow Cytometry

Journal: Journal of Nanobiotechnology

Article Title: Orchestrated Cu 2+ -coordinated tetracycline-porphyrin self-assembly remodels tumor microenvironment for photo-enhanced immuno-chemodynamic therapy

doi: 10.1186/s12951-025-03486-9

Figure Lengend Snippet: In vivo immunomodulatory effects of LP/CuTT. Representative flow cytometric analysis and quantification of tumor-infiltrating M1 TAMs gated on CD11b + and CD80 + cells ( A , B ), CD4 + T cells gated on CD3 + and CD4 + cells ( C , D ), and CD8 + T cells gated on CD3 + and CD8a + cells ( E , F ) from the upper right quadrant of the scatter diagrams in tumors after 17 days treatments. G - L ) Representative flow cytometric analysis and quantification of M1 TAMs, CD4 + T cells and CD8 + T cells in spleen after different treatments. Serum levels of cytokines M ) TNF-𝛼, N ) IL-1β, and O ) IL-10 in mice after different treatments detected by ELISA. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001, compared to control. # p < 0.05, ns indicates no significant difference

Article Snippet: PE anti-mouse CD206/MMR antibody, FITC anti-mouse CD86 antibody, APC anti-mouse CD80 antibody, FITC anti-mouse CD3 antibody, PerCP anti-mouse CD8 antibody, and APC anti-mouse CD4 antibody were purchased from Elabscience (Wuhan, China).

Techniques: In Vivo, Enzyme-linked Immunosorbent Assay, Control