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. 2013 Mar 28;8(3):e60165. doi: 10.1371/journal.pone.0060165

Figure 1. KLF11 expression in human urogenital tissues, uterine eutopic endometrium and endometriosis.

Figure 1

(A) Pooled Human Organ-specific RNA (1 µg/reaction) was analyzed for KLF11 mRNA expression by PCR. mRNA levels of the housekeeping gene GAPDH were simultaneously assessed as loading control. Relative organ-specific KLF11 mRNA expression levels were determined by densitometric comparison to corresponding GAPDH levels. KLF11 mRNA expression levels were increased in urogenital tissues such as the kidney, ovary, placenta, testes and uterus (Figure 1A, arrows) compared to other non-urogenital tissues. (B) Expression of KLF11 mRNA and protein were also assessed in two cell lines Ishikawa, a well-differentiated endometrial adenocarcinoma cell line and Human Endometrial Stromal Cells (T-HESC), which are frequently used as model endometrial epithelial and stromal cell lines respectively. KLF11/KLF11 were expressed in both cell-lines; GAPDH/α-TUBULIN were used as a reference controls. To qualitatively demonstrate KLF11 expression in each of these endometrial cell lines, lysate for RNA or protein extraction was obtained from one 10-cm cell culture dish of confluent cells. Due to much lower abundance of stromal compared to epithelial cells there appears to be differential KLF11/KLF11 expression between the cell-types. However, this difference was also reflected in expression of the loading controls GAPDH/α-TUBULIN between the two cell lines. (C, D) KLF11 expression was also evaluated by immunohistochemistry in eutopic endometrium as well as in ectopic endometrial implants (magnification: 200×: panel; 400×: inset). KLF11 was expressed in the nuclei and cytoplasm of epithelial as well as stromal cells in both eutopic endometrium as well as in endometriotic implants. KLF11 expression was diminished in endometriotic implants compared to eutopic endometrium. Representative samples shown. (E) KLF11 expression levels were compared and scored in paired eutopic endometrium and ectopic endometriotic implants obtained from the same patient (N = 28 paired samples). The expression was significantly reduced in the implants (epithelium: 103±2.6; stroma: 40.4±3.9) compared to that in eutopic uterine endometrium (epithelium: 278±2.5; stroma: 114±8.7). * and. = p<0.05 and represent comparisons between epithelial and stromal cells in eutopic and ectopic endometrium respectively.