Journal: Molecular medicine reports

Article Title: BCG stimulation promotes dendritic cell proliferation and expression of VDR and CYP27B1 in vitamin D‑deficient mice.

doi: 10.3892/mmr.2019.10780

Figure Lengend Snippet: Figure 2. Effect of BCG on BMDC differentiation and maturation. (A) Cell morphology during BMDC induction from days 1 to 6, as observed by light micros- copy. (B) BCG stimulated BMDC maturation. (C) Phenotypic changes of the BMDCs induced by BCG stimulation and (D) the levels of the surface molecules of DCs. All images were captured under identical magnification and microscopy conditions. **P<0.01 vs. 0 mg/ml in the normal control or vitamin D‑deficiency groups. ##P<0.01, vitamin D‑deficiency group vs. normal control group. Scale bar, 20 µm. BCG, Bacillus Calmette‑Guérin; BMDC, bone marrow‑derived dendritic cell; CD11c, integrin alpha‑X; CD80, T‑lymphocyte activation antigen CD80; MHC‑I, major histocompatibility complex class I; MHC‑II, major histocompatibility complex class II; CD86, T‑lymphocyte activation antigen CD86.

Article Snippet: The cell concentration was adjusted to 1x106 cells/ml and the cells were fixed with 0.1 ml polyformaldehyde for 30 min at 25 ̊C Subsequently, fluorescein isothiocyanate‐labeled integrin alpha‐X (CD11c; 1:50; 130‐110‐700; Miltenyi Biotech), T‐lymphocyte activation antigen CD86 (CD86; 1:50; 130‐102‐506; Miltenyi Biotech), T-lymphocyte activation antigen cd80 (cd80; 1:50; 130‐116‐459; Miltenyi Biotech), major histocompatibility complex class i (MHc-i; 1:50; ab95572; abcam) and MHC‐II (1:50; 130‐112‐386; Miltenyi Biotech) were added, respectively.

Techniques: Microscopy, Control, Activation Assay, Immunopeptidomics

Journal: iScience

Article Title: Microglial transcription profiles in mouse and human are driven by APOE4 and sex.

doi: 10.1016/j.isci.2021.103238

Figure Lengend Snippet: Figure 2. Sex and APOE genotype drive amyloid pathology and microglial gene expression profiles in APOE-FAD mice (A) Representative images of immunohistochemical staining for amyloid-b (red) and the microglial marker IBA1 (green) in male and female APOE3- and APOE4-FAD mice. Scale bar, 50 mm. (B) Samples separate by APOE and sex along PC1, with APOE3-FAD males clustering separately from mice of both sexes and APOE3-FAD females. A large proportion of the genes driving this separation (76.4%) are DAM-APOE genes. APOE3-FAD males are shown in yellow, APOE3-FAD females in gray, APOE4-FAD males in magenta, and APOE4-FAD females in teal. (C) Heatmap of the DAM-APOE genes associated with sample clustering along PC1. Gene expression levels were normalized, scaled, and centered and are displayed as Z scores. (D and E) Levels of the DAM-APOE marker, CD11c, are increased in APOE3-FAD females and in APOE4-FAD mice of both sexes, both at the (D) transcript and (E) protein levels. (D) Gene expression of Itgax in normalized counts; data are presented as mean (GSEM) values; n = 5–7/group. *p < 0.05, **p < 0.01; one-way ANOVA with Tukey’s multiple comparisons test correction. (E) Representative immunofluorescence images of microglia (IBA1, red) and CD11c (green) in the hippocampal subiculum of male and female APOE3- and APOE4-FAD mice. Scale bar, 25 mm.

Article Snippet: For IBA1/CD11c double labeling, antigen retrieval was performed using boiling EDTA and sections were incubated for 2 days at 4 C in primary antibodies directed against IBA1 (1:500) and CD11c (#NB110-97871, 1:500, Novus Biologicals).

Techniques: Gene Expression, Immunohistochemical staining, Staining, Marker

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 induction of M2 TAMs, the 1-6/TAM spheroids formulation, the transwell co-incubation, and their respective treatments. b Western blot analysis of GPX4 and FSP1 expression in M2 TAMs after different treatments. The experiment was repeated three times independently with similar results. Uncropped blots in Source Data. c Quantification of grayscale intensity of GPX4 and FSP1 in ( b ) ( n = 3 independent experiments). Protein expression levels in PBS group were normalized to 1. d Live/dead staining of M2 TAMs after different treatments (Scale bar = 200 μm). Viable cells were stained with calcein-AM (green), and dead cells were stained with PI (red). The experiment was repeated three times independently with similar results. e Quantification of the percentage of viable and dead M2 TAMs in ( d ) ( n = 3 independent experiments). f Flow cytometric analysis of Annexin V-FITC/propidium iodide (PI)-stained M2 TAMs (gated on F4/80 + CD206 + macrophages) after different treatments. g Quantification of the percentage of apoptotic cells in ( f ) ( n = 3 independent replicates). The experiment was repeated twice independently with similar results. h The cell proliferation of M2 TAMs after different treatments measured by CCK8 ( n = 3 independent experiments). The concentrations of SF and vF contained in nanoparticles were both 3 μM. i Fluorescence images of C11 BODIPY 581/591 -stained M2 TAMs after different treatments (Scale bar = 100 μm). The experiment was repeated three times independently with similar results. j Fluorescence images of live/dead stained 1-6/TAM spheroids after different treatments (Scale bar = 500 μm). Viable cells were stained with calcein-AM (green), and dead cells were stained with PI (red). Z-stack scanning of the spheroids was performed with slices distanced by 14.36 μm. The experiment was repeated three times independently with similar results. k Flow cytometric analysis of anti-CRT-stained Hepa1-6 cells after different treatments. l Quantification of the percentage of CRT positive cells in ( k ) ( n = 3 independent replicates). The experiment was repeated twice independently with similar results. m The ATP assay of Hepa1-6 cells after different treatments ( n = 3 independent experiments). n The HMGB1 assay of Hepa1-6 cells after different treatments ( n = 3 independent experiments). o Flow cytometric analysis of anti-CD80/CD86-stained BMDCs (gated on CD11c + DCs) after different treatments. p Quantification of the percentage of mature DCs in ( o ) ( n = 3 independent replicates). The experiment was repeated twice independently with similar results. q The assay of TNF, IL-6, and IL-12 in BMDCs after different treatments ( n = 3 independent experiments). Unless specified otherwise, error bars represent the mean ± SEM. Statistical significance was determined by one-way ANOVA with Tukey’s test ( c , g , h , l – n , p , q ) and P -values were indicated. Source data are provided as a Source Data file. The elements in Fig. 5a 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: Formulation, Incubation, Western Blot, Expressing, Staining, Fluorescence, ATP Assay

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 Body weight of mice with intravenous treatments during a 28-day period ( n = 5 independent mice). b The blood analysis for the liver/kidney functions including ALT, AST, BUN, and CRE was determined on day 28 following the treatments ( n = 5 independent mice). c The curve of injected drug concentration (ID %) versus time point was plotted in subcutaneous HCC mice ( n = 6 independent mice). d Fluorescence imaging of the biodistribution in orthotopic HCC mice at 12 h after different treatments (tumors marked with yellow circles; n = 6 independent mice). e Tumor-to-background ratio for different treatments in ( d ) ( n = 6 independent mice). f Quantitative biodistribution analysis of DiR-labeled nanoparticles in major organs and liver tumors in ( d ) ( n = 6 independent mice). g Fluorescence images of Sv@PM-M2p inside HCC cells, M2 TAMs, M1 TAMs, DCs, and T cells in tumors ( n = 6 independent mice; Scale bar = 100 μm). Sv@PM-M2p was labeled with Rhodamine (red), the nuclei and cell markers (HCC, CK8; M2, CD206; M1, CD86; DC, CD11c; T, CD3) were stained with DAPI (blue) and antibody (green), respectively. h Quantification of the percentage of colocated cells in ( g ) ( n = 6 independent mice). Unless specified otherwise, error bars represent the mean ± SEM. Statistical significance was determined by one-way ANOVA with Tukey’s test ( c , e , f , h ) and P -values were indicated. Source data are provided as a Source Data file.

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: Injection, Concentration Assay, Fluorescence, Imaging, Labeling, 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 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 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