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. 2020 Sep 22;12(9):2718. doi: 10.3390/cancers12092718

Figure 3.

Figure 3

Chemokine-mediated migration of preadipocytes and TAMs. (A) Culture media (CM) from LNCaP cells stably transfected with SFMBT2 shRNA promote the migration of 3T3-L1 preadipocytes. However, the addition of antibodies against CXCL8, CCL2, CXCL10, and CCL20, and a mixture of antibodies reduces the migration of 3T3-L1 preadipocytes. After incubation of 3T3-L1 preadipocytes with antibodies against indicated chemokines in culture media from LNCaP cells stably transfected with SFMBT2 (shSFMBT2) or control shRNA (shCont), transwell assay was performed. Migrating 3T3-L1 preadipocytes were counted. (B) Treatment of CXCL8, CCL2, CXCL10, and CCL20 promotes the migration of 3T3-L1 preadipocytes. The addition of antibodies against CXCL8, CCL2, CXCL10, and CCL20 reduces the migration of 3T3-L1 preadipocytes. After incubation of 3T3-L1 preadipocytes with antibodies against indicated chemokines in RPMI 1640 media, transwell assay was performed. Migrating 3T3-L1 preadipocytes were counted. (C) Increased expression of CCR2, CCR4, CCR6, CXCR1, CXCR2, and CXCR3 in 3T3-L1 preadipocytes incubated with culture media from LNCaP cells stably transfected with SFMBT2 shRNA. RNA was analyzed. (D) Culture media from LNCaP cells stably transfected with SFMBT2 shRNA promote the migration of TAMs. However, the addition of antibodies against CXCL8, CCL2, CXCL10, and CCL20, and a mixture of antibodies reduces the migration of TAMs. After incubation of TAMs polarized from Raw264.7 cells with antibodies against indicated chemokines in culture media from LNCaP cells stably transfected with SFMBT2 or control shRNA, transwell assay was performed. Migrating TAMs were counted. (E) Treatment of CXCL8, CCL2, CXCL10, and CCL20 promotes the migration of TAMs. However, the addition of antibodies against CXCL8, CCL2, CXCL10, and CCL20 reduces the migration of TAMs. After incubation of TAMs polarized from Raw264.7 cells with antibodies against indicated chemokines in RPMI 1640 media, transwell assay was performed. Migrating TAMs were counted. (F) Increased expression of CCR2, CCR6, CXCR1, CXCR2, and CXCR3 in TAMs polarized from Raw264.7 cells incubated with culture media from LNCaP cells stably transfected with SFMBT2 shRNA. RNA was analyzed. All data represent mean ± S.E.M. Significance values were * p ≤ 0.05, ** p ≤ 0.01, and *** p ≤ 0.005.