Supplemental Material: Necrostatin-1 Attenuates Skeletal Muscle Injury Following Limb Ischemia-Reperfusion Injury

The supplementary information provides additional biochemical, functional, histological, inflammatory, and molecular data supporting the findings reported in the main manuscript. Included are plasma creatine kinase (CK) and lactate dehydrogenase (LDH) measurements across multiple reperfusion time points; longitudinal grip-strength assessments; individual animal-level histopathological injury scores and the corresponding H&E scoring rubric; full cytokine-array membranes, a cytokine-expression heatmap, and complete normalized cytokine-array results with statistical comparisons; additional Western blot data for necroptosis- and cell-death-associated proteins, including RIPK1, phosphorylated RIPK1, phosphorylated MLKL, NF-κB, and cleaved caspase-8; analysis of IDO1 protein abundance following Nec-1 treatment; whole-section p-MLKL immunofluorescence images; and antibody and dilution information used for immunoblotting and immunofluorescence analyses.

Together, these materials provide supporting data and methodological detail for the assessment of skeletal muscle injury and Nec-1-associated effects following L-IRI.</p>