Mi se

After 3 h of Eh infection, colons of Math1^GFP mice were surgically removed, imaged ex vivo using an in vivo Xtreme 4MP-imaging platform (Bruker, Billerica, MA, USA) to detect GFP expression. The imaging was performed in two steps. The first one is reflectance imaging (2 s exposure time) and the second one was with excitation at 470 nm and emission at 535 nm (5 s exposure time) i.e., fluorescent imaging. Images were acquired and analyzed from the in vivo Xtreme using Bruker molecular imaging software MI SE (version 7.1.3.20550). GFP expression in the colon was quantified by measuring the mean fluorescence (after background subtraction) in a constant ROI.

Animal experiments were performed using protocols approved by the Ethics Committee of Zhujiang Hospital of Southern Medical University. All experiments followed the regulatory standards. Female 6- to 8-week-old NOD/SCID IL-2RγCnull (NSG) mice were purchased from Biocytogen. Mice were bred under specific pathogen-free conditions and randomized into 4 groups: (1) control, (2) gilteritinib, (3) FLT3scFv/NKG2D-CAR T cell and (4) combination of FLT3scFv/NKG2D-CAR T cell and gilteritinib. On day zero, luciferase-transfected MOLM-13 cells (2 × 10⁶ cells/ml) were injected via the tail vein of all mice. Starting on day 7, gilteritinib was dissolved in 4% DMSO and then administered by intraperitoneal injection at a dose of 15 mg/kg 5 days per week for 3 weeks in Groups 2 and 4. From day 8, a single dose of FLT3scFv/NKG2D-CAR T cells (2.5 × 10⁶ cells) based on the published reported [25 (link)] was injected via the tail vein in Groups 3 and 4. Leukaemia progression was monitored by serial bioluminescence (BL) imaging using an IVIS Lumina imaging system (PerkinElmer, USA) following D-luciferin substrate administration (0.3 mg/g body weight, i.p.) (Promega, USA). Data were analysed using Bruker MI SE (Bruker FX Pro, USA). Survival curves were generated.

Bacteria translocation during DSS and Citrobacter infection was performed by homogenizing tissue lysates in sterile PBS and normalizing protein content between samples. Serial dilutions were then plated on MacConkey agar and CFU enumerated the following day. For Citrobacter quantification, MacConkey agar plates contained 100 μg/mL streptomycin. For whole body imaging of bioluminescent Citrobacter, animals were anaesthetized with 2% isofluorane carried in 2% O₂, kept constant at 37 °C via air circulation and imaged on their ventral side on days 3, 5, 7, 10, and 14 using an in vivo Xtreme 4MP imaging platform (Bruker, Billerica, MA, USA). The imaging protocol contained three steps: reflectance imaging (2 s exposure time), bioluminescent imaging (10 s exposure time) and an additional Xray imaging step (10 s exposure time). Binning was kept constant at 4 × 4. Images from the in vivo Xtreme were acquired and analyzed using Bruker molecular imaging software MI SE (version 7.1.3.20550, Bruker, Billerica, MA, USA). Citrobacter-associated bioluminescence expression in the abdomen in Vamp8^+/+ and Vamp8^-/- littermates was quantified by measuring the mean bioluminescence (after background subtraction) in a constant region of interest (ROI), which was kept constant over the time period of imaging.