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. 2016 Mar 10;7(15):20869–20889. doi: 10.18632/oncotarget.8029

Figure 4. Hypoxic/necrotic core in GemOE tumors enhances Ac-HMGB1 secretion.

Figure 4

(A–C) Adjacent sections from a GemOE tumor IHC stained for geminin (A), c-Abl (B) and HMGB1 (C). (D) The level of circulating HMGB1 measured using specific ELISA assay performed on serum isolated from samples collected 7 weeks after mice were injected in mammary fat pads with naïve HME cells (n = 10, no tumors developed) or GemOE cells (n = 30, tumor-bearing, p = 0.00042). Two different sets (E, G, I and K) and (F, H, J and L) of adjacent sections from GemOE orthotopic mammary tumors stained with H & E (E and F), or IHC stained for geminin (G and H), HMGB1 (I and J) as well as HIF-1α (K) or hypoxyprobe (L). N denotes necrosis within these tumors that are shown adjacent to the hypoxic cells as indicated by high HIF-1α or hypoxyprobe staining. These cells are also expressing cytoplasmic HMGB1. (M) The levels of HMGB1 detected using specific ELISA assay released from iGem9, iG197, iG240 or iG257 cells grown under normoxic (N) or hypoxic (H) conditions. Experiments were done in triplicates 3 different times, **represents p < 0.001. (N) The level of acetylated HMGB1 passively diffused from naïve HME, iG197, iG240 or iG257 cells after repeated freeze and thaw cycles.