Journal: bioRxiv

Article Title: Macropinosomes are a site of HIV-1 entry into primary CD4 + T cells

doi: 10.1101/2025.03.07.642068

Figure Lengend Snippet: A macropinocytosis inhibitor inhibits HIV-1 fusion with primary CD4 + T cells. (A) A schematic illustration of the experimental procedure performed for panels B-J. Activated primary CD4 + T cells and A3.01 cells were pretreated with either EIPA or T-20 or left untreated for 15 minutes at 37°C and inoculated with HIV-1 containing BlaM-Vpr for 2 hours at 37°C. Inoculated cells were loaded with CCF2-AM at 15°C for 1 hour and incubated overnight at room temperature. The cells were then labeled with either an anti-CXCR4-APC/Cy7 or CCR5-APC/Cy7 antibody and analyzed by flow cytometry. (B) Representative flow plots of surface expression of CXCR4 on primary CD4 + T cells. (C and D) Quantification of the relative density of surface CXCR4 on primary CD4 + T cells (C) and A3.01 cells (D). (E) Representative flow plots of the percentages of cleaved CCF2 + cells in CXCR4 + cells are shown. (F and G) Quantification of the relative percentages of cleaved CCF2 + cells (fusion efficiency) in CXCR4 + primary CD4 + T cells (F) and A3.01 cells (G). (H) Quantification of the relative density of surface CCR5 on primary CD4 + T cells. (I) Representative flow plots with the percentages of cleaved CCF2 + cells in CCR5 + cells are shown. (J) Quantification of the relative percentages of cleaved CCF2 + cells (fusion efficiency) in CCR5 + primary CD4 + T cells. Data are presented as mean ±SD in C, D, F-H, and J. The experiments were performed three times with primary CD4 + T cells isolated from three donors in B, C, E, F, and H-J. The experiments were performed three times with A3.01 cells independently in D and G. The p values were determined using Dunnett’s test following one-way ANOVA in C, E-G. *, p <0.05; **, p <0.01; ***, p <0.001; ****, p <0.0001; n.s., not significant.

Article Snippet: Cells were washed with 1xPBS(-) once and labeled with anti-CD3-APC/Cy7 or CD3-AlexaFluor647 (1:100; clone UCHT1, BioLegend) and CD4-AlexaFlor647 or CD4-FITC (1:100; clone OKT4, BioLegend) antibodies in FACS buffer for 30 minutes at 4°C.

Techniques: Incubation, Labeling, Flow Cytometry, Expressing, Isolation

Journal: Nature Communications

Article Title: Quasi Fe MIL-53 nanozyme inducing ferroptosis and immunogenic cell death for cancer immunotherapy

doi: 10.1038/s41467-025-57542-x

Figure Lengend Snippet: a Schematic illustration of experimental process of in vivo antitumor immunity. b Flow cytometry analysis of mature DCs of tumors after saline and Q-MIL-53(Fe) treatment. c Flow cytometry analysis of CD4 + T cells and CD8 + T cells of tumors after saline and Q-MIL-53(Fe) treatment (the CD4 + T cells and CD8 + T cells were gated on CD3 + T cells). d, e IF staining of tumor sections for Granzyme B ( d ) and FOXP3 ( e ) expression. Images are representative of three biologically independent mice. Scale bar = 200 μm. f Schematic illustration of the experimental process of in vivo immunotherapeutic effects. g The tumor growth curves of 4T1 tumor-bearing mice after indicated treatments. The significance between each of the multiple groups was calculated using one-way ANOVA. h Images of the harvested tumors. i The recorded weight of tumors after different treatments. The significance between each of the multiple groups was calculated using one-way ANOVA. j The recorded weight of 4T1 tumor-bearing mice after indicated treatments. k Photographs of tumors and major organs (heart, liver, spleen, lung, and kidney). l H&E staining of the tumors sections. Scale bar = 100 μm. Images are representative of three biologically independent mice. All data are presented as means ± SD ( n = 5 biologically independent animals for g , i , j ). Source data are provided as a Source Data file.

Article Snippet: Antibodies (anti-GAPDH, Beyotime, 1:1000; Goat anti-Rabbit IgG (H + L) Secondary Antibody, HRP, Beyotime, 1:1000; Goat anti-Mouse IgG (H + L) Secondary Antibody, HRP, Beyotime,1:1000; GPX4 Rabbit Polyclonal Antibody, Beyotime, 1:1000; PD-L1 Rabbit Monoclonal Antibody, Beyotime, 1:1000; Calreticulin Rabbit Monoclonal Antibody, Beyotime, 1:500; HMGB1 Rabbit Polyclonal Antibody, Beyotime, 1:200; IgG (H + L) (Alexa Fluor® 488-conjugated Donkey Anti-Goat IgG (H + L), Servicebio, 1:500; IgG (H + L), Cy3 conjugated Donkey Anti-Goat IgG (H + L), Servicebio, 1:500; FITC Anti-CD80 Rabbit pAb, Servicebio, 1:100; APC Anti-CD3 antibody, abcam, 1:100; PE Anti-CD86 antibody, abcam, 1:100; PE Anti-CD8 alpha antibody, abcam, 1:100; FITC Anti-CD4 antibody, abcam, 1:100.) were purchased from corresponding commercial sources with specific dilutions.

Techniques: In Vivo, Flow Cytometry, Saline, Staining, Expressing