Fig 5. MLF and DnaJ-1 control crystal cell differentiation.

(A) MA-plot of DESeq2 results for RNAseq data comparison between control and mlf^-/- lz>GFP⁺ blood cells sorted by FACS from third instar larvae. Genes that are significantly upregulated or downregulated in the mlf mutant (adjusted p-value<0.01) are highlighted in red or blue, respectively. Red triangles: genes with log₂ fold change >5. (B) Pie chart showing the number of expressed genes in lz>GFP⁺ cells and the number of upregulated (red) or downregulated (blue) genes in the mlf mutant. (C) Heat map of “crystal cell”-associated genes differentially expressed (p-value<0.01) between control and mlf mutant lz>GFP⁺ cells. Differential gene expression as per comparison to the mean of the 6 samples (ctr 1, 2, 3 and mlf 1, 2, 3) is displayed as log₂ scale. Hierarchical clustering was performed using R-Bioconductor. (D-O) Immunostainings against GFP and in situ hybridization against CG7860 (D-F), Oscillin (G-I), Jafrac1 (J-L) and CG6733 (M-O) in blood cells from lz-GAL4,UAS-mCD8-GFP/+ control (D, G, J, M), mlf^-/- (E, H, K, N) or dnaj-1^-/- (F,I, L,O) third instar larvae. RNA expression only is shown in the lower panels. Nuclei were stained with Topro3. Scale bar: 10 μm.