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. 2011 Apr 6;286(21):19047–19064. doi: 10.1074/jbc.M110.213983

FIGURE 4.

FIGURE 4.

Cdx4 expression was increased by HoxA10 overexpression and decreased by HoxA10 knockdown in U937 myeloid leukemia cells. A, HoxA10 expression level influenced Cdx4 mRNA abundance in a dose-dependent manner. U937 cells were stably transfected with a vector to overexpress HoxA10 or Cdx4 (or empty expression vector) and a vector to express HoxA10-specific shRNAs, Cdx4-specific shRNAs, or scrambled shRNA controls. Expression of HoxA10 and Cdx4 mRNA was determined by real-time PCR. * and **, statistically significant differences in HoxA10 expression with HoxA10 overexpression or knockdown, respectively (p < 0.00001, n = 6). *** and #, statistically significant differences in HoxA10 expression with Cdx4 overexpression (p < 0.02, n = 6). ##, significant increase in HoxA10 expression in shCdx4-expressing cells with HoxA10 overexpression (p < 0.0001, n = 6). ### and &, significant differences in Cdx4 expression in cells with HoxA10 overexpression or knockdown (p < 0.0001, n = 6). && and &&&, significant increase in Cdx4 expression with Cdx4 overexpression (p < 0.0001, n = 6). +, significant increase in Cdx4 expression in cells with shHoxA10 upon Cdx4 overexpression (p < 0.0001, n = 6). B, HoxA10 expression level influenced Cdx4 protein abundance. U937 cells were stably co-transfected with a vector to overexpress HoxA10 or Cdx4 (or empty expression vector) and with a vector to express HoxA10-specific shRNAs or bp-swapping, scrambled shRNAs. Total cell lysates were separated by SDS-PAGE, and Western blots were serially probed with antibodies to Cdx4, HoxA10, and tubulin (as a loading control). C, HoxA10 overexpression partly rescued Cdx4 protein expression in cells with Cdx4 knockdown. U937 cells were stably co-transfected with a vector to overexpress HoxA10 (or expression vector control) and a vector to express Cdx4-specific shRNAs or scrambled shRNAs. Total cell lysates were separated by SDS-PAGE, and Western blots were serially probed with antibodies to Cdx4, HoxA10, and GAPDH (as a loading control). Error bars, S.E.