Extended Data Fig. 4: Relationship of MYC multimers to nucleoli and PML bodies.
From: MYC multimers shield stalled replication forks from RNA polymerase
a. Immunofluorescence of MAX and MYC upon treatment with DOX (1 µg/ml, 24 h) and MG-132 (20 µM, 4 h) in U2OSMYC-Tet-On cells. Scale bar: 5 µm (n = 3). b. Immunofluorescence of MAX and MYC upon treatment with DOX (1 µg/ml, 24 h), MG-132 (20 µM, 4 h) and 10058-F4 (150 µM, 6 h) in U2OSMYC-Tet-On cells. Scale bar: 5 µm (n = 3). c. Bar plot quantifying results from (b). Shown are fold changes of numbers of multimers compared to MG-132 treated cells. The number of cells analyzed are: DMSO (n = 494), MG-132 (n = 825), 10058-F4 (n = 1087), MG-132+10058-F4 (n = 852) over three independent experiments. Data are shown as mean ± S.E.M. Two sided Mann-Whitney U test was used for comparison to MG-132 treated cells with following p-values: DMSO < 2.2e-16, MG-132+10058-F4 < 2.2e-16. d. dSTORM of MYC using antibody Y69 in U2OS cells treated MG-132 (20 µM), CBL0137 (5 µM) and PlaB (1 µM) for 4 h or HS (42 °C for 30 min), at saturated labeling conditions (n = 3). Scale bar: 5 µm. e. Confocal immunofluorescence using RNA Polymerase I (RNAPI), PML (PCC = 0.465) and MYC in U2OSMYC-Tet-On cells treated with DOX (1 µg/ml, 24 h) and MG-132 (20 µM, 4 h). Scale bar: 5 µm (n = 3). f. Immunofluorescence of MYC and Fibrillarin upon treatment with DOX (1 µg/ml, 24 h), MG-132 (20 µM, 4 h) or CX5461 (0.5 µM, 4 h) in U2OSMYC-Tet-On cells. Scale bar: 5 µm (n = 3). In above panels, unless indicated otherwise, n indicates the number of independent biological replicates.
