Fig. 2: Als3 uptake is required for inducing immune responses.
From: Fungal Als proteins hijack host death effector domains to promote inflammasome signaling
a BMDMs were incubated with Alexa Fluor 488-conjugated Als3 or Als3S170Y at either 4 °C (top panels) for 1 h or 37 °C for 15 or 30 min (middle and bottom panels), fixed, and then observed under confocal microscopy. Samples were stained with an antibody against the endosome marker EEA1. At least 100 cells were checked in each experiment and the pictures are representative. Scale bars, 10 μm. b BMDCs were stimulated with Als3 (15 μg ml−1) for 24 h in the absence or presence of DMSO, Pitstop 2 (25 μM), or R406 (1 µM). IL-1β and TNF levels were measured by ELISA. Data are mean ± s.e.m. of two independent experiments. P-values were calculated with an unpaired two-tailed t test (n = 6). c BMDMs were either unstimulated (medium) or stimulated with C. albicans hyphae in the absence or presence of Pitstop 2. IL-1β cleavage and caspase-1 activation were analyzed by immunoblotting. Data in (a, c) are representative of three independent experiments. At least 18 randomly fields were checked for the results in (a, b).
