Fig. 2: Elevated expression of MGAT1 inhibits tumor immune response in 2D and 3D in vitro coculture systems.

a Schematic diagram of the coculture assays. Created in BioRender. Zhu, Y. (2025) https://BioRender.com/v30i433. b The proliferation of MDA-MB468, MDA-MB468-MGAT1-OE, and MDA-MB468-MGAT1-KD cells (green) in 2D coculture with pre-activated PBMCs was monitored by time-lapse image-based quantification. The data are presented as mean ± SEM from three replicates from a representative experiment. c MDA-MB468, MDA-MB468-MGAT1-OE, and MDA-MB468-MGAT1-KD, and MDA-MB231, MDA-MB231-MGAT1-OE, and MDA-MB231-MGAT1-KD cells were cocultured with pre-activated human PBMCs, and IFNγ⁺, TNFα⁺, Granzyme B⁺, Ki-67⁺ CD8⁺ T cell populations were measured and quantified using flow cytometry. d The infiltration of PBMCs (red) in 3D spheroids constructed with MDA-MB468, MDA-MB468-MGAT1-OE, and MDA-MB468-MGAT1-KD or MDA-MB231, MDA-MB231-MGAT1-OE, and MDA-MB231-MGAT1-KD was visualized with Z-stack imaging using a Lionheart microscope. e The IFNγ⁺, TNFα⁺, Granzyme B⁺, and Ki-67⁺ CD8⁺ T cell populations from 3D coculture with MDA-MB468, MDA-MB468-MGAT1-OE, and MDA-MB468-MGAT1-KD cells were measured and quantified using flow cytometry. f The proliferation of MGAT1-WT/OE/KD MDA-MB468 cells (green) in 2D coculture with pre-activated purified CD8⁺ T cells was monitored by time-lapse image-based quantification. The data are presented as mean ± SEM from three replicates from a representative experiment. g The IFNγ⁺, TNFα⁺, and Ki-67⁺ CD8⁺ T cell populations from 2D coculture with MDA-MB468, MDA-MB468-MGAT1-OE, and MDA-MB468-MGAT1-KD cells were measured and quantified using flow cytometry. Data (represented as means ± SEM), images, and flow cytometry are representative of three independent experiments. Statistical significance was determined using one-way ANOVA with Tukey’s multiple comparisons tests (c, e, g) or two-way ANOVAs followed by Tukey’s multiple comparison tests (b, f). Source data are provided as a Source Data file.