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. 2019 Feb 12;10(2):133. doi: 10.3390/genes10020133

Figure 3.

Figure 3

Addition of the Flag-Sly transgenes to males with NPYq deficiency rescues SLY1 and SLY2 expression deficiency. (A–D) Western blot was performed with whole testes lysates obtained from XY males and from males with moderate NPYq deficiency (2/3NPYq-) with (tsgic) and without (neg sib) Flag-Sly transgene addition. Two transgenic lines were tested: 16D positive for the Sly1 transgene (A,B) and 6P positive for both the Sly1 and Sly2 transgenes (C,D). Levels of protein expression shown in panels (A,C) were quantified with ImageJ software, normalized to a non-specific band (B,D) with XY data serving as the normal expression baseline. The normalization for line 6P was also done to Ponceau signal (Figure S7). The data represent an average ± SEM with n = 3. Statistical significance (t-test, p < 0.05): a different from XY; b different from neg sib. (E) Western blot was performed with cytoplasmic (C) and nuclear (N) protein lysates from the whole testes obtained from XY males and males with severe NPYq deficiency (9/10NPYq-) with (tsgic) and without (neg sib) Sly transgene addition. Two transgenic lines were tested: 30A positive for the Flag-Sly1 transgene and 6P carrying both the Sly1 and Sly2 transgenes. Due to the scarcity of testicular material, the reference gene was not included in this analysis.