Skip to main content
Fig. 6 | Genome Biology

Fig. 6

From: The Tug1 lncRNA locus is essential for male fertility

Fig. 6

Overexpression of TUG1-BOAT compromises mitochondrial membrane potential. a Construct and transfection scheme. Human and mouse ORF1, and mouse Tug1 cDNA lacking the ORF1 region (Tug1 cDNA ∆mORF1) were inserted into pcDNA3.1(+). Chloromethyl-X-rosamine (CMXR) was added to visualize the mitochondria 48 h post-transfection. After staining, cells were fixed and processed for anti-FLAG immunofluorescence (IF) or Tug1 RNA FISH. b Maximum intensity projections of z-stacks acquired 48 h post-transfection of 3T3 cells with indicated plasmids and staining with CMXR. Tug1 RNA overexpression was monitored by Tug1 single-molecule RNA FISH (gray) and TUG1-BOAT by immunostaining against the FLAG tag (green). CMXR is shown in red and DAPI in blue. On the right, quantification of cells positive for Tug1 RNA or TUG1-BOAT and mitochondria by CMXR (n = 50). Scale bar is 5 μm. c Maximum intensity projections of z-stacks acquired 48 h post-transfection of 3T3 cells with the indicated plasmids, stained with CMXR (red) and immunostained against mitochondrial membrane translocase TOM20 (gray). On the right, quantification of cells overexpressing TUG1-BOAT and lacking CMXR staining showing an intact mitochondrial membrane assessed by TOM20. The nuclei were stained with DAPI (blue). Scale bar is 5 μm

Back to article page