Expression of lola isoforms in the Drosophila brain. (a-d) RNA in situ analysis of lola isoforms. (a1) An antisense probe generated against the common region of lola labels uniformly throughout the brain at 0 h APF. (a2) A sense control to the same probe shows little specific staining. (a3) A magnified view of the AL reveals lola expression in all PN cell bodies. PN cell bodies are marked by white arrows, while dotted while lines demark the rough area of the AL neuropil in each section that is not stained by DAPI. Midline to the left, lateral to the right. Isoform specific probes to isoform L (b), isoform Q (c) and isoform T (d) show different patterns of expression throughout the brain at 0 h APF, while sense control probes (b2-d2) show little specific labeling. Closer inspection of AL regions at a higher magnification (b3-d3) reveals that most isoforms appear to be expressed in PNs. Scale bars: 20 μm (a3-d3); 200 μm (a1-d2). DIG-labeled RNA probe in red, DAPI in blue, GFP in green. See Additional file 2 for in situ analysis of additional lola isoforms and additional labeling of section morphology. (e) Quantitative RT-PCR of laser-captured PN enriched-samples verifies in situ results that most isoforms are expressed in PNs. Additionally, different lola isoforms are expressed at different levels at 0 h APF, with about 100-fold difference between highest and lowest expression levels. Data are displayed by lola isoform on the X-axis and by relative abundance on a log scale on the Y-axis, where relative abundance has been determined against the level of actin42 expression. Error bars represent the standard deviation from four independent samples, and each sample was tested independently five times per device. Horizontal solid line represents a confidence limit of the average relative expression of samples near the detection limit based on CT values and reproducibility.