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. 2017 Aug 3;8(46):80545–80559. doi: 10.18632/oncotarget.19849

Figure 5. Inhibiting UPR signaling components in the tumor epithelial cell affects macrophage polarity.

Figure 5

(A) Control, IRE1, PERK, or GRP78 transfected 4T1B breast cancer cells were plated in an ACEA E-plate and RAW 264.7 macrophages were added to the E-plate. Each well was treated with 1 μg/mL LPS and the cell index was measured by electrical impedance. n = 3; *p < 0.05. (B) Conditioned media from control or GRP78 siRNA transfected 4T1B or ZR-75-1 breast cancer cells were used to treat RAW 264.7 cells for 24 hours. iNOS and Arg-1 were measured by Western blot hybridization. Protein loading was normalized to β-actin. n = 3; *p < 0.05. (C) Vehicle treated, LPS treated, ZR-75-1 conditioned media (CM), or GRP78-silenced ZR-75-1 conditioned media were used to treat RAW 264.7 macrophage cells for 24 hours. Macrophages were stained for CD68-FITC (green), CD80-Cy5 (red), or CD206-cy7 (pink) and counterstained with DAPI. The M1/M2-like macrophage population was determined by immunocytochemistry. (D) Vehicle treated, LPS treated, ZR-75-1 conditioned media, or GRP78-silenced ZR-75-1 conditioned media were used to treat RAW 264.7 macrophage cells for 24 hours. Macrophages were stained for IL12-Cy3 (yellow) or IL10-cy7 (pink) and counterstained with DAPI. The M1/M2-like macrophage population was determined by immunocytochemistry.