Fig. 5

Attenuation of TCDD-induced spreading of actin stress fibers by Jdp2 deficiency. A Cells were serum starved overnight and then treated with DMSO and TCDD for 24. At the end of treatment, the cells were rinsed with PBS, fixed with 4% formaldehyde, and processed for F-actin staining with Alexa 488-conjugated phalloidin. Nuclei stained blue with DAPI. For each treatment, at least five fields were acquired. B The fluorescence intensity was measured in WT and Jdp2^–/– MEFs in the presence or absence of 50 nM TCDD. CCCTC binding factor CTCF and cell area were measured as described in the “Methods” section. C Extension of cell spreading of WT and Jdp2^−/− MEFs after exposure to TCDD, and p-MLC2 and F-actin expression upon TCDD challenge in WT and Jdp2^−/− MEFs. Representative staining of F-actin and p-MLC2, and merged images are shown. D, E Western blot analysis of p-MLC2 and total MLC2 expression (D) along with the quantitative results (E). Cropped figures are shown. See Supplementary Figure S18 for the original full-length blot images. The intensity of each band was then quantified. The relative value was normalized to β-Actin and shown as ratio. E for cells harvested at 6 h after treatment with 0–200 nM TCDD. The cell lysates were processed to measure p-MLC2 content, as described in the “Methods” section. Data are presented as mean ± SEM from three independent experiments and p values were obtained