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. 2023 Oct 16;13(1):69–82. doi: 10.1093/stcltm/szad069

Figure 3.

Figure 3.

In vitro kidney organoid coculture assay with T cells. (A) Schematic of the in vitro coculture experiment and readouts. (B–C) Surface expression on organoid cells of (B) HLA-ABC and (C) CD54 (ICAM) measured by flow cytometry. Kidney organoid cells cultured without the addition of T cells (unstimulated) were used as a control and set to 1 for comparison. Results are shown of kidney organoids cultured with IFN-γ, CMV-reactive T cells (CMV), or HLA-A2-reactive T cells (Allo-A2) (n = 2 independent experiments for IFN-γ, n = 3 for others). (D) CD137 expression on CD8+ T cells cultured with control or B2M–/– kidney organoids measured by flow cytometry. T cells cultured alone (unstimulated) were used as a control and set to 1 for comparison (n = 3 independent experiments). (E) IFN-γ secretion in the supernatant by the T cells cultured with control or B2M–/– kidney organoids measured by ELISA. T cells cultured alone (unstimulated) were used as control for spontaneous IFN-γ secretion (n = 3 independent experiments). B2M–/– organoid clones are indicated with separate symbols; #2 (circle), #7 (triangle), and #10 (star). Results are shown as mean ± SD and significance was evaluated using one-way ANOVA with Dunnett’s correction for multiple testing, comparing each sample to the unstimulated control (***P < .001, ****P < .0001).