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. 2020 Apr 3;10(4):543. doi: 10.3390/biom10040543

Figure 4.

Figure 4

AnaR cells recruit and activate macrophages through leptin signaling. (a) Quantitative real-time PCR for mRNA expression of ARG1 (Arginase 1), IL10 (Interleukin-10), markers of pro-tumor macrophages and IL12 (Interleukin-12), INOS (inducible Nitric Oxide Synthase), markers of tumor-inhibitory macrophages, in RAW 264.7 cells after incubation with conditioned medium (CM) derived from MCF-7 aro or AnaR cells. (b) Transmigration assay in RAW 264.7 cells using as a chemoattractant charcoal-stripped serum media (-) or conditioned medium (CM) derived from MCF-7 aro or from AnaR cells. (c) Transmigration assay in RAW 264.7 cells treated or not with the peptide LDFI (1 µM) using as chemoattractant AnaR-CM or AnaR-CM in which leptin was immunodepleted by incubation with a mouse monoclonal specific antibody against leptin (LepAb). (d) Transmigration assay in RAW 264.7 cells treated with vehicle (-), leptin (1000 ng/mL) alone or in the presence of the peptide LDFI (1 µM) using as a chemoattractant MCF-7 aro CM. * p < 0.05; ** p < 0.005; *** p < 0.0005.