Figure 1
PARP12 is a trans-Golgi localized protein. (a) Representative confocal microscopy images of HeLa cells untreated (a,b) or treated with Nocodazole (33 μM, 3 h) (c,d), fixed and labeled with anti-PARP12 (red), anti-GM130 (a cis-Golgi marker, green) and anti-Golgin-97 (a TGN-Golgi marker, blue) antibodies. (a) Zoom: enlarged view of Golgi area. Quantification of Manders’ colocalization coefficient (MCC) of PARP12/GM130, PARP12/Golgin-97 and GM130/Golgin-97 (as internal control) are shown in the graph. Data are mean ± SD from n = 20. Statistical significance was calculated by unpaired Student’s t-test; ***p < 0.001. (b) Zoom 1 and 2: higher magnification images of Golgi area shown in a. (c) Clear separation of GM130 (green) and Golgin-97 (blue) markers in ministacks under Nocodazole treatment. Zoom: enlarged view of ministacks. Far right graph: quantification of Manders’ colocalization coefficient (MCC) under Nocodazole treatment. Data are mean ± SD from n = 20. Statistical significance was calculated by unpaired Student’s t-test; ***p < 0.001. (d) Zoom 1–6: enlarged view of single isolated mini stacks numbered in c. White arrow across the stack was used for line-scan analyses and the fluorescence intensity distribution of markers along this line was quantified and normalized to their respective peak values. The images shown are representative of three independent experiments. Scale bars, 10 μm.
