Axiocam mrm rev 2 ccd camera

Immunostaining against SPD-2 (“969LA”, 1:800) was performed as previously described [4 (link)]. A Zeiss Axio Imager Z1 microscope equipped with an Axiocam MRm REV 2 CCD camera was used for capturing pictures. Immunostainings against PSer5 of Pol II CTD (“3E8”, Millipore, 1:25) and PSer2 of Pol II CTD (“3E10”, Millipore, 1:25) were performed as previously described [44 (link)]. DNA was visualized using 4-6-Diamidino-2-phenylindole (DAPI). Pictures were taken using an Axio Imager M2 microscope and a Yokogawa CSU W1 Duel camera. Images were processed with Fiji and Adobe Photoshop in an identical manner and imported into Adobe Illustrator.

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.