X-linked gene expression levels for each independent sample type.

(A) qRT-PCR analysis of RNA levels for five X-linked genes in DR3#6; LctraIR7 or wild type XX and XY mid-staged pupae. Expression of X- linked gene is higher in DR3#6; LctraIR7 XX than XY pupae for all genes except Lczfh2 (Lcaru, P = 0.008; LcCG1909, P = 0.007; LcGW, P = 0.005; LcMav, P = 0.007; LcZfh2, P = 0.071; Student’s t-test). Expression of X-linked genes is higher in wild type XY pupae than XX pupae for two of the five genes (Lcmav, P = 0.025; LcCG1909, P = 0.039; Student’s t-test) Transcript levels were normalized to 28S rRNA. Expression levels were averaged over three biological replicates (DR3#6 LctraIR7) or 4 biological replicates (wild type) and standard error shown. Significance denoted on graph as * P < 0.05 or ** P < 0.01. (B) qRT-PCR analysis of RNA levels for five X-linked genes in LctraKI heterozygous or homozygous XX and XY hemi-sected adults. Expression of X-linked genes in LctraKI homozygous XX compared to XY is higher in two out of the five genes tested (Lcaru, P = 0.074; LcCG1909, P = 0.015; LcGW, P = 0.029; LcMav, P = 0.701; LcZfh2, P = 0.536; Student’s t-test). No significance was observed comparing LctraKI heterozygous XX and XY adults, however LcGW expression was visibly increased in LctraKI heterozygous XX flies (P > 0.05; LcGW, P = 0.183; Student’s t-test). Transcript levels were normalized to 28S rRNA. Expression levels were averaged over four biological replicates and standard error shown. Significance denoted on graph as * P < 0.05.

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