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Ex vivo cultures combined with vivo-morpholino induced gene knockdown provide a system to assess the role of WT1 and GATA4 during gonad differentiation

Fig 6

Single- and double-knockdown of WT1 and GATA4 affect mRNA levels of hedgehog pathway components in XY and XX gonads.

XY (A-C) and XX (D-F) gonads, dissected from murine embryos (12.5 dpc), were incubated for 72 h with Wt1 and/ or Gata4 vivo-morpholinos as described in Fig 4. Relative transcript levels were determined by qRT-PCR and normalized to Sdha transcripts according to the 2ΔΔCT method [34]. Results are shown as fold differences between cultures treated with mismatch vs. Wt1- (A, D) and mismatch vs. Gata4-morpholino (B, E), respectively. Panels C and F represents the effect of double-knockdown of WT1 and GATA4. Error bars represent S.E.M. from independent biological replicates (n ≥ 8). scale = log2. Statistical differences are indicated by asterisks: *p<0.05, **p<0.01, ***p<0.005, ****p<0.001, paired t-test.

Fig 6