1. Role of microRNA-21 and programmed cell death 4 in the pathogenesis of human uterine leiomyomas 
J. Browning Fitzgerald, B.S., Vargheese Chennathukuzhi, Ph.D., Faezeh Koohestani, M.S., Romana A. Nowak, Ph.D., Lane K. Christenson, Ph.D. 
Fertility and Sterility 
Volume 98, Issue 3, Pages e2 (September 2012)
DOI: /j.fertnstert
Copyright © 2012 American Society for Reproductive Medicine Terms and Conditions

2. Figure 1
Differential expression of programmed cell death 4 (PDCD-4) and microRNA-21 (miR-21) in leiomyoma and myometrial tissues and immortalized cell lines, UtLM (leiomyoma) and UtM (myometrial). (A) miR-21 levels in myometrial and leiomyoma tissues (n = 12, P=.08). (B) Western blot analysis of PDCD-4 isoforms in three representative paired leiomyoma and normal myometrial samples of a total of 11 examined are shown. (C) Western blot analysis of PDCD-4 isoforms in UtLM and UtM cell lines. *Mean ± standard error of the mean (n = 3) for the 51 kd band between cell types are different (P<.05). (D) qRT-PCR analysis of miR-21 and PDCD-4 mRNA levels in UtLM and UtM cell lines. *Mean ± standard error of the mean (n = 3) are different (P<.05) between UtLM cells and UtM cells.
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3. Figure 2
MicroRNA-21 (miR-21) regulation of programmed cell death 4 (PDCD-4) in immortalized myometrial (UtM) and leiomyoma (UtLM) cells. Locked nucleic acid (LNA) oligonucleotides complementary to miR-21 (LNA-miR-21) or a scrambled (LNA-scr) control were transfected into (A) UtM cells or (B) UtLM cells. *Mean ± standard error of the mean (n = 3) PDCD-4 (51 kd isoform) levels normalized to actin are different (P<.05) between LNA-scramble and LNA-miR-21 treated cells.
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Copyright © 2012 American Society for Reproductive Medicine Terms and Conditions

4. Figure 3
Knockdown of microRNA-21 (miR-21) in UtM and UtLM cells increases cleaved caspase-3. Locked nucleic acid (LNA) oligonucleotides complementary to miR-21 (LNA-miR-21) or a scrambled (LNA-scramble) control were transfected into immortalized (A) myometrial (UtM) cells or (B) leiomyoma (UtLM) cells. Western blot analysis showed a significant induction of cleaved caspase-3 in the LNA-miR-21 treated UtM and UtLM cells across three independent experiments.
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5. Figure 4
Expression of phosphorylated elongation factor-2 (EF-2) in immortalized myometrial (UtM) and leiomyoma (UtLM) cells after knockdown of microRNA-21 (miR-21). (A) Basal ph-EF2 expression levels in UtM cells and UtLM cells. *Mean ± standard error of the mean (n = 3) ph-EF2 levels normalized to actin are different (P<.05). (B) Locked nucleic acid (LNA) oligonucleotides complementary to miR-21 (LNA-miR-21) or a scrambled (LNA-scr) control were transfected into UtM cells or UtLM cells. *Mean ± standard error of the mean (n = 3) ph-EF2 levels normalized to actin are different (P<.05) between LNA-scramble and LNA-miR-21 treated cells.
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6. Supplementary Figure 1
PDCD-4 expression across paired normal and leiomyoma tissue using polyclonal anti-PDCD-4 antibody from (A) Sigma and (B) ProSci. P represents individual patients.
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Copyright © 2012 American Society for Reproductive Medicine Terms and Conditions

7. Supplementary Figure 2
(A) Efficacy of miR-21 knockdown after LNA-21 treatment in UtM and UtLM cells. (B) PDCD-4 mRNA levels after LNA-NS (scrambled) and LNA-21 treatment in UtM and UtLM cells. ∗∗Mean ± standard error of the mean (n = 3) miR-21 levels normalized to U6 are different (P<.01) between LNA-miR-21 (LNA-21) and LNA-scrambled (LNA-NS).
Fertility and Sterility  , e2DOI: ( /j.fertnstert )
Copyright © 2012 American Society for Reproductive Medicine Terms and Conditions