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HRP-conjugated secondary antibodies (rabbit IgG: 111035144, mouse IgG: 115035003, 1:3000 dilution in 3% BSA, Jackson ImmunoResearch) and the ECL-2 (Thermo Scientific) or ECL (GE Amersham) chemiluminescent reagent were used to visualize the bands using a Bio-Rad ChemiDoc imaging system running Image Lab V4.0 software

Classically activated M1 macrophages adopt a glycolytic metabolic profile characterized by rapid glucose uptake, enhanced lactate production, and interruption of the tricarboxylic acid (TCA) cycle at the citrate and succinate nodes

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Methods Cell lines, cell culture, and reagents All cell lines were obtained from ATCC, except murine PC cells (KPC K8484: Kras G12D/+

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Collectively, these findings strengthen the rationale for developing multifaceted strategies that leverage intestinal microbiota modulation to mitigate DKD progression (231)