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. 2014 Oct 15;289(49):34065–34073. doi: 10.1074/jbc.M114.604041

FIGURE 3.

FIGURE 3.

CRABP2 inhibits mammary carcinoma cell growth by two distinct mechanisms. A, M-2−/− cells stably expressing EGFP (Ctrl), EGFP-CRABP2 (2), or EGFP-2ΔNLS (2ΔNLS). Immunoblotting demonstrates similar expression of WT and mutant CRABP2. GAPDH was used as a loading control. B, cells were cultured in delipidated medium for 48 h, treated with vehicle or RA (200 nm) for 4 days, and counted. C, M-2−/− cells (3 × 106) that stably overexpress EGFP (Ctrl), EGFP-CRABP2 (2), or EGFP-CRABP2ΔNLS (2ΔNLS) were injected subcutaneously into female NCrnu/nu mice, and tumor growth was monitored. Data are mean ± S.E. (mice: control, n = 20; CRABP2 and CRABP2ΔNLS, n = 10 each). *, p ≤ 0.05 versus control; ‡, p ≤ 0.05 using two-tailed Student's t test. D–H, levels of denoted mRNAs (D and F) and proteins (E, G, and H) in tumors expressing EGFP (Ctrl), CRABP2 (2), or CRABP2ΔNLS (2ΔNLS). D and F, levels of mRNAs assessed by qPCR. Data are mean ± S.E. (n = 3–5). *, p ≤ 0.05 versus control using two-tailed Student's t test. E, G, and H, left, immunoblots using denoted antibodies. Right, quantification of immunoblots. Data are mean ± S.E. (E and G, n = 8; H, n = 4). *, p ≤ 0.05 versus control using two-tailed Student's t test. Error bars represent S.E.