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Fig. 6 | Breast Cancer Research

Fig. 6

From: A functional role for the cancer disparity-linked genes, CRYβB2 and CRYβB2P1, in the promotion of breast cancer

Fig. 6

CRYβB2 overexpression significantly increases IL6 secretion and enhances immune cell chemoattraction. a qRT-PCR analysis of IL-6 mRNA levels in the indicated cell models +/- CRYβB2 overexpression. b IL-6 ELISA. Cells were grown to 80% confluence, washed, then incubated in serum-free media 24 h. Conditioned media collected, concentrated, corrected for total protein then analyzed. Representative model of all three triple-negative breast cancer models. c The indicated SUM model was grown to 80% confluence on the bottom of a migration chamber, washed, then incubated in serum-free media 24 h, then U937 cells were plated in serum-free media on the top chamber and allowed to migrate towards the indicated cell model for 4 h. Data are the number of cells migrated per field counted, right panels are representative crystal violet-stained images of migrated cells. d Representative immunoblots of SUM159 overexpressing CRYβB2 cells +/- the addition of 75 ng/ml recombinant human IL-6 receptor blocking antibody. e Proliferation of SUM159 cells overexpressing CRYβB2 ± 75 ng/ml recombinant human IL-6 receptor blocking antibody, or the indicated cell line treated with the blocking antibody (f). Cells were plated at 25,000 cells per well in triplicate and counted every 24 h for 96 h. Blocking antibody was added each day. All data represent mean ± SE of a minimum of three independent experiments. *p < 0.01. #p = 0.052. 159, SUM159; +C, CRYβB2 overexpression; +P1, CRYβB2P1 overexpression; P-/-, CRYβB2P1 knockout; C-/-, CRYβB2 knockout; +C+P, CRYβB2 and CRYβB2P1 dual overexpression

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