Fig 1: Assessing the effect of M42 protein treatment on viability of hiPSC derived macrophages. Human iPSC macrophages were treated with HTRA1 (A), DAG1 ectodomain (B), TMEFF2 ectodomain (C), MDK (D), GPC5 (E), SDC4 ectodomain (F), SFRP1 (G), SPON1 (H), and PTN (I) at 3 µg/mL, 1 µg/mL, and 100 ng/mL for 24 h. Viability was assessed and expressed as relative luminescence units (RLU). N = 2 or 3 independent biological experiments (one independent macrophage factory set‐up; three technical replicates/biological repeat). Error bars correspond to mean ± standard deviation (SD). Repeated measures one‐way ANOVA followed by Dunnett's multiple comparisons test.
Fig 2: Assessing the effect of M42 recombinant proteins on phagocytosis of apoptotic SH‐SY5Y cells in human induced pluripotent stem cell (hiPSC)‐derived macrophages. (A) Summary graph showing fold change in total phagocytosis assessed over 24 h relative to control (vehicle‐treated cells) across all tested proteins at three concentrations: 3 µg/mL, 1 µg/mL, and 100 ng/mL. (B–F) Phagocytosis following treatment with HTRA1 (B), DAG1 ectodomain (C), TMEFF2 ectodomain (D), MDK (E), and GPC5 (F). Left panels show comparison between phagocytosis levels in vehicle‐ and protein‐treated hiPSC macrophages quantified by area under the curve (AUC). Right panels show a representative trace from one biological repeat (n = 3 technical replicates). Levels of phagocytosis are measured as total integrated intensity of pHrodo‐labeled apoptotic neurons normalized to the confluency of macrophage layer (TII/confluency). Cytochalasin D (CytoD, 10 µM) treatment was included in every experiment as a negative control. N = 3 to 4 independent biological experiments (one or two independent macrophage factory set‐ups; three technical replicates/biological repeat). Error bars correspond to mean ± SD. Repeated measures one‐way ANOVA followed by Dunnett's multiple comparisons test. *p < 0.05; **p < 0.01.
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