Histofix

Cells used for morphological analysis were filled with an intracellular solution containing 0.1–0.5% biocytin (Sigma Aldrich, USA) through the patch-pipette while recording. Acute slices were fixed in 4% Histofix (Carl Roth, Germany) after recording. 2–10 days later, slices were washed in 1× PBS (phosphate buffered saline), permeabilized in 0.2% PBST (0.2% Triton in 1× PBS) and stained overnight with a Streptavidin-coupled Alexa594-conjugated antibody (life technologies, USA). Slices from 5xFAD mice were additionally stained with an Alexa488-coupled 6E10 Antibody (Covance, USA) for Aβ plaque staining. After washing in 1× PBS, slices were mounted in ProLong Gold Antifade (life technologies, USA).
Neurons were imaged with a fixed-stage Leica TCS SP5 II microscope (Leica, Germany) and the Leica LAS AF Lite Software (Leica, Germany). Z-stacks from whole neurons were imaged with a 40× oil-immersion objective (Leica, Germany) with the following parameters: voxel size x/y = 0.758 μm, z = 0.209 μm. Z-stacks from dendrites were taken with a 63× oil-immersion objective (Leica, Germany) with the following parameters: voxel size x/y = 0.08 μm, z = 0.168 μm.
Z-stacks of DG granule cells were semi-automatically traced with Neuronstudio (CNIC, Mount Sinai School of Medicine, USA) and Sholl analysis was performed. Dendritic spines were also counted semi-automatically with Neuronstudio.

Superficial and deep CA1 pyramidal neurons used for morphological analysis were filled with a solution containing 0.1–0.5% biocytin (Sigma‐Aldrich) through the patch pipette while recording. Acute slices were fixed in 4% Histofix (Carl Roth) after recording. After 2–10 days, the slices were washed in 1× PBS (phosphate‐buffered saline) for 3× 10 min. Permeabilization was performed for 1 h in 0.2% PBST (0.2% Triton X‐100 in 1× PBS). Slices were stained overnight with Alexa 594‐conjugated Streptavidin directed against biocytin (Life Technologies). On the next day, the slices were washed again for 3× 10 min in 1× PBS. After air‐drying the slices at RT for 1 h, they were mounted with a coverslip in ProLong Gold Antifade (Life Technologies).

All animal experiments were conducted in compliance with European and German regulations. Protocols were approved by the responsible Federal State authority and ethics committee (Thüringer Landesamt für Verbraucherschutz, permit number: 03-006/09). Female BALB/c mice (Charles River, Germany) weighing 18–20 g were housed in groups of five in individually ventilated cages with free access to water and food. For infection, C. albicans was grown for 12 h at 30°C in YPD medium, washed three times in sterile PBS, and diluted to the desired concentrations. The infection dose was confirmed by plating. On day 0, the mice were infected via the lateral tail vein with 2.5 × 10⁴C. albicans cfu/g body weight. The health status of the mice was examined at least twice daily. Liver was collected 6 and 24 h post-infection, fixed in 10% neutral buffered formalin (Histofix, Carl Roth, Karlsruhe, Germany), embedded in paraffin, and sectioned at 4 µm thickness.

BENOs were fixed with 4% formaldehyde solution (Histofix, Carlroth) for 2 h at 4 °C. Subsequently, they were washed twice with PBS and blocked for 30 min at 4 °C with staining buffer (StB; 5% FBS, 1% BSA, 0,5% Triton X-100 in PBS). BENOs were incubated with primary antibodies diluted in StB for 2 days at 4 °C (100 µl/BENO). Upon washing with StB for 6–8 h, BENOs were incubated with secondary antibodies and Hoechst 33342 (Sigma) for another 2 days at 4 °C. After StB washings for a total of 6–8 h BENOs were mounted on glass coverslips. An antibody list with respective dilutions is provided in Supplementary Data 2. WmIF was visualized using confocal imaging performed on a Zeiss LSM 710 confocal microscope equipped with ZEN 2010 software.

Whole lung tissue sections were fixed in Histofix (4.5%, Carl Roth) overnight and embedded in paraffin. Slices (4 μm) were stained with either H&E or PAS and examined using a Zeiss Axio Imager.M2 with the AxioCam MRc camera. Tissue inflammation and infiltration was evaluated on H&E stained sections. PAS-positive goblet cells were quantified in percent of counted cells which have been determined by visible nuclei.