CEST MRI data (both before and after injection) were extracted using custom-written MATLAB functions to calculate the Z-spectra of mean pre-injection images. For the analysis of the results, the mean pre-injection Z-spectra were subtracted from all post-injection images. The “smooth” function in MATLAB was used to apply a moving average filter at each frequency. This moving average filter was applied on the time series data to remove fluctuations in signal due to motion. The smoothening factor used in the averaging process was ~40 neighboring images, with this choice based on the size of the data and the range of fluctuations. After smoothening, 10 to 20 images were averaged to generate the corresponding parameter maps (contrast, FF, and pH). Images acquired between 40 and 50 min after injection were considered for the calculation of parametric maps on Mut+/− and Mut−/− mice. For calculating the contrast and FF, CEST images at 4.2 ppm were used. Time-contrast uptake curves were obtained by calculating the average ST over pixels selected by two ROIs, one over each kidney. Maximum contrast maps were generated by calculating the pixel-by-pixel maximum ST on two ROIs, one over each kidney, overlaid on the corresponding T2W image. The time-averaged maximum contrast images were calculated by taking 10 to 20 images at corresponding post-injection time. The FF was calculated by determining the percentage of pixels in the kidneys with a contrast >20% of the maximum contrast detected at 4.2 ppm for each mouse. The time-averaged FF over 10 images was used to generate the FF bar plot.

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