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The micro-PET images of the BDL and CCl₄ models of liver fibrosis and control mice (sham group in 21 d, n=9; control group in 12 wk, n=10) were obtained using a micro-PET scanner (Siemens) as reported.¹² Briefly, static PET/CT images were acquired 1 hour after i.v. tail vein injection of [¹⁸F]AlF-ND-bisFAPI (4.44–5.55 MBq, 0.1 mL), with an acquisition of 10 minutes. Mice were anesthetized and scanned in the prone position. Blocking experiments were performed in the BDL model mice by coinjection of the FAP competitor DOTA-FAPI-04 (50 μg/mouse, n=5) at day 21 after operation. Images were reconstructed using a 3-dimensional ordered-subset expectation maximum algorithm (Siemens). Attenuation correction was performed using the unenhanced low-dose CT data. To quantify the tracer uptake in liver sections, the regions of interest were drawn manually on decay-corrected whole-body coronal images using the Inevon Research Workplace 4.1 software (Siemens). The mean standardized uptake value for each region of interest was recorded to avoid the influence of weight change.

For dynamic micro-PET imaging studies, either ¹⁸F- or ⁶⁸Ga-labeled tracer (5.55–11.1 MBq, n = 3) were injected through the lateral tail vein into mice bearing A549-FAP tumor xenograft using an Inveon Micro-PET/CT scanner (Siemens; Erlangen, Germany). Image studies were conducted using a three-dimensional ordered-subset expectation maximum (OSEM) algorithm (Siemens, Erlangen, Germany). A549-FAP tumor-bearing mice were co-injected for the blocking study using DOTA-FAPI-04 (100 nmol/mouse, n = 3) as a competitor. The images and regions of interest (ROIs) were produced using Inevon Research Workplace 4.1 software (Siemens, Erlangen, Germany).