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Fig. 5 | BMC Genomics

Fig. 5

From: Segmental duplications and evolutionary acquisition of UV damage response in the SPATA31 gene family of primates and humans

Fig. 5

Expression analysis of SPATA31 RNA and protein within human primary fibroblast (HFF) cells. a RT-PCR analysis on cDNA isolated from total human RNA (Clontech) and from Mouse RNA. The PCR primers were designed to amplify the highly conserved region within the long coding exon for human. For mouse, we amplified a region spanning exons 1–3. The UBE1 gene was used as control. Quantitative PCR for the same tissue samples is shown in Additional File 8. b Antibody staining of HFF cells with fixation under dark conditions versus a treatment with 200 J/m2 UVC and 24 h further growth at two different magnifications (63x and 100x). Nuclei in the first column are stained with DAPI (blue), and SPATA31 staining is shown in the second column (red). In the third column, the green staining in the 63x pictures is cellular cytoskeleton detected with an antibody against tubulin and in the 100x columns the nucleolus with an antibody against Per2 [45]. See Additional File 12 for further quantification of SPATA31 response of re-localization upon different exposure to UVC. We noted some variation of relative SPATA31 protein localization between cytoplasm and the nucleus depending on the cell cycle and fixation protocol as well as dark–light conditions during fixation similar to other UV response proteins such as H2Ax (reviewed in [46] and see Methods)

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