Panel A shows fluorescence images of DMSO- and DOX-treated cells stained for DDR protein and GM130, with yellow arrowheads indicating protein localization. Panel B shows dot plots of Logarithm base 10 of Golgi intensity (normalized to control) for Trans-Golgi and Cis-Golgi proteins after Double-strand break damage (DOX) across Base excision repair, Microhomology-mediated end joining, Mismatch repair, Cell division and DNA damage response, Translesion synthesis, Homologous recombination, Prefoldin complex, and Cell cycle groups. Panel C shows dot plots of Logarithm base 10 of Golgi intensity (normalized to control) after Double-strand break damage (DOX) comparing Trans-Golgi and Cis-Golgi protein localization across the indicated protein groups. Panel D shows dot plots of Logarithm base 10 of Total Golgi plus nuclear intensity (normalized to control) after Double-strand break damage (DOX) for Trans-Golgi and Cis-Golgi proteins across the indicated functional categories.
DOX-induced redistribution, related to Fig. 2. (A) Representative images of HeLa-K cells stained with antibodies against DDR proteins and the Golgi marker, GM130. Cells were treated with DOX (40 μM, 3 h), prior to fixation. Yellow arrowheads denote Golgi membranes. Scale bars, 10 μm. (B–D) Quantification of DDR protein intensity changes upon DOX treatment. Relative intensity of DDR proteins (B) at the Golgi and (C) in the nucleus and (D) total Golgi and nuclear signal normalized to untreated control. Data represent the mean ± SEM (n = 3 biologically independent experiments with at least 200 cells analyzed for each protein in control and treatment conditions). The proteins are classified according to their Golgi localization patterns (Fig. 2 E). Proteins shown in orange co-localize with the trans-Golgi marker TGN46, while those in blue co-localize with the cis-Golgi marker GM130. Proteins whose localization showed no clear preference for either marker are shown in grey. Statistical significance was determined using a two-tailed unpaired Student’s t test; ns, *P < 0.05, **P < 0.01, ***P < 0.001, and ****P < 0.0001, compared with untreated control.