Fig. 6

The expression of Nvd, Spok, Dib and Sad is compromised in the dfr¹⁴ mutant. a Ecdysone biosynthesis steps in the Drosophila prothoracic gland. Steroidogenic enzymes Nvd, Sro, Phm, Dib, and Sad are sequentially required to convert cholesterol to ecdysone. b Immunostaining of control (upper panels), homozygous dfr¹⁴ (middle panels), and hemizygous dfr¹⁴ /Df (lower panels) BRGCs for the steroidogenic enzymes Nvd, Sro, Phm, Dib, and Sad. The immunostaining of the enzymes is shown in red, DAPI staining in blue. Scale bars 50 μm, except for the images stained for Sad (100 μm in these images). c Quantification of relative fluorescence intensity in b. The expression of Nvd, Dib, and Sad was significantly reduced in dfr¹⁴ and dfr¹⁴ /Df, compared to control. d RT-qPCR results of the steroidogenic genes. Transcript levels of nvd, spok, and dib were significantly decreased in dfr¹⁴ (n = 5; Q* < 0.05, Q** < 0.01). e Ecdysone concentrations were quantified in a 2-, 4-, or 12-h time window from early to late L3. n = 4 for each time point. For comparison, 40 h ALE is approximately equal to 88 h ALH in Fig. 3k, l. Asterisks denote significant differences at each time point (n = 4; Q*** < 0.0001)