GFP Quantification in Caenorhabditis elegans Germline

Reporter GFP quantifications were done as described (Seelk et al., 2016 (link)). Briefly, fluorescent micrographs were recorded with Zeiss Axio Imager Z1 microscope and a Zeiss Axiocam MRm REV 2 CCD camera was used to capture images. For each germline (n=19 in control RNAi, n=23 in prp-19(RNAi) and n=19 in Fig. S2), three nuclei from the distal-most zone were taken and intensities quantified using Fiji. Then GFP intensities were normalized to the picture background and corrected with the average autofluorescence measured in wild-type (N2) gonads at the corresponding temperatures. Images subject to direct comparison were taken at identical exposure times and were processed with Adobe Photoshop CS5.1 in an identical manner.