Fig. 2.

PIEZO1 orchestrates a unique immune response in human iMGLs. A Toxicity as count of Cytotox Green cells per confluence for iMGLs treated with 2–20 µM Yoda1 and live-imaged for 60 h. Normalized to positive control (PC) of 200 µM 1-Methyl-4-phenylpyridinium iodide. N = 3 in n = 3. B Quantification at 48 h. C Representative images of confluency and labeling for fluorescent Cytotox Green reagent. D–O The cells were treated with 5 µM Yoda1 24 h before starting the assays. D Respective toxicity for iMGLs treated with 5.0 µM GsMTx4 and E quantification at 48 h. n = 6 wells. F Spare respiratory capacity calculated from the oxygen consumption rate (OCR) profiles in mitostress test. n = 7 with N = 5. G Ratio of OCR to extracellular acidification (ECAR). H A mitostress energy map depicting OCR and ECAR compared to 20 ng/ml LPS. I Scratch wound density normalized to vehicle. Areas under the curves (AUC) presented as bars. N = 3 in n = 2. Green fluorescence intensity of phagocytosed. J pHrodo beads (n = 4 with N = 3) and K 0.5 µM green HiLyte Aβ 1–42 (N = 3 in n = 1) per confluence over 5 h with AUC normalized to vehicle. L Lysosomal activity over time as intensity of pH-sensitive fluorescent red LysoView 540 reagent per confluency. n = 5 with N = 3. Representative images of M scratch wounds at 24 h overlayed with masks for the original scratches at 0 h and at 24 h, N cells with internalized green fluorescent pHrodo beads that show as black dots outside the cells at 3 h; O cells with internalized Hilyte Aβ42 488 at 5 h; and, P cells treated with 20 µM Yoda1 and fluorescent red LysoView 540 reagent. Two-way ANOVA with Sidak’s multiple comparisons or unpaired t-test. Significance ***p < 0.001, **p < 0.01, *p < 0.05. Data as mean ± SEM. All data repeated in n = experiments/batches with N biological replicates. See also Additional file 1: Fig. S2