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. 2024 Jun 7;10(23):eadj3194. doi: 10.1126/sciadv.adj3194

Fig. 8. Caudal IKKβCA-CM increases macrophage migration and polarization toward an inflammatory phenotype.

Fig. 8.

(A) Study design schematic of whole-organ in vitro culture and CM collection from caudal control (AcanCre−/−;Ikk2cafl/fl) and IKKβCA IVDs harvested 1 week after recombination. (B) Protein concentrations (pg/ml) within CM analyzed after 1, 3, and 5 days in culture. UD, undetectable reading. Letters (a), (b), and (c) indicate statistically significant (P < 0.05) different groupings (n = 3 per genotype). (C) Quantification of M0, M1, and M2 macrophage migration through a transwell membrane via DAPI nuclear count (n = 3). (D) Representative images of IF staining for CD38 and CD206 within BMDMs cultured in 2D monolayer and stimulated with control or IKKβCA CM. DAPI nuclear stain (blue). Scale bars, 100 μm. (E) Quantification of % positivity for CD38 and CD206 within bone marrow–derived macrophages treated with CM (n = 3). (F) Gene expression of M1 and M2 phenotypic markers within M0, M1, or M2 macrophages in basal media, or M0 macrophages with or without IVD CM stimulation (n = 3). Letters (a), (b), and (c) indicate statistically significant (P < 0.05) different groupings.