INHA protein levels in SCs.

<p>Inha levels in SCs transfected with plasmids pshRNA-1, pshRNA-2, pshRNA-3, and pshRNA-negative 48 h after transfection. Lanes 1 to 4 represent the pshRNA-1, pshRNA-2, pshRNA-3, and pshRNA-negative plasmids, respectively. The normalized ratio for INHA was calculated by dividing the mean signal intensity for 3 biological replicates by the mean signal intensity with ACTB. Data are presented as the mean ± SEM, with different letters (a, b) was significant (<i>P</i><0.05), as evaluated using Student's paired t test.</p

Effects of <i>Inha</i> silencing on the cell cycle in SCs.^*

<p>*Each experiment was repeated 3 times. Values represent the mean ± SEM (n = 3 in each group).</p><p>**<i>P</i><0.01, as evaluated by Student's paired t test.</p

RNAi recombinant plasmids expressed in SCs.

<p>GFP expression in SCs with (<b>B</b>) or without (<b>A</b>) transfection of RNAi recombinant plasmids 48 h after transfection. The scale bar represents 500 µm.</p

INHA profiles.

<p>INHA in male neonate mice on days 1, 7, 10, 14, 21, 28, 35, and 56 post partum. The normalized ratio for INHA protein was calculated by dividing the mean signal intensity from 3 biological replicates by the mean signal intensity with ACTB.</p

Relative mRNA expression level of Inha, Inhba and Inhbb.

<p>Relative mRNA expression level of Inha, Inhba and Inhbb from male neonates mice at days 1, 7, 10, 14, 21, 28, 35, and 56 of age.</p><p>Values presented by mean ± S.E.M, n = 4 in each age group. In the horizontal row, significance level was p<0.05 between values scripted with different little letters, and not significant with same little letters. In the vertical row, significance level was p<0.05 between values scripted with different big letters, and not significant with same big letters.</p

<i>Inha</i> mRNA expression in transfected cells.

<p>The <i>Inha</i> mRNA levels in SCs transfected with RNAi recombinant plasmids pshRNA-1, pshRNA-2, pshRNA-3, or pshRNA-negative 48 h after transfection. Data are presented as the mean ± SEM (n = 3 in each group). For bars with different letters (a, d; b, d; c, d), the difference was significant (<i>P</i><0.01).</p

Restriction mapping and sequencing analysis.

<p>Restriction mapping analysis shows (<b>A</b>) the pshRNA-1 (Lane 1), pshRNA-2 (Lane 2), pshRNA-3 (Lane 3), and pshRNA-negative (Lane 4), RNAi-Ready pSIREN-RetroQ-ZsGreen Vector (Lane 5) plasmids and sequencing of plasmids pshRNA-1 (<b>B</b>), pshRNA-2 (<b>C</b>), and pshRNA-3 (<b>D</b>).</p

Protein levels in transfected SCs.

<p>INHBA, INHBB, Cyclin D1, Cyclin E, and P21 levels in SCs transfected with the pshRNA-2 and pshRNA-negative plasmids 48 h after transfection. The normalized ratio for each protein was calculated by dividing the mean signal intensity from 3 biological replicates by the mean signal intensity with ACTB. Data are presented as the mean ± SEM. *P<0.05, as evaluated using Student's paired t test.</p

Target sequences of mouse inhibin alpha (<i>Inha</i>).

<p>Target sequences of mouse inhibin alpha (<i>Inha</i>).</p

Sequences of primer pairs for quantitative real-time PCR.

<p>Sequences of primer pairs for quantitative real-time PCR.</p