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Fig. 8 | Biological Research

Fig. 8

From: In situ isolation of nuclei or nuclear proteins from adherent cells: a simple, effective method with less cytoplasmic contamination

Fig. 8

Validation of the effectiveness and sensitivity of in situ nuclear isolation for the evaluation of pP65 by western blotting. a Quantification of total nuclear proteins of HUVECs obtained via different methods. The final volume of the lysates is approximately 100 μL for all groups. b Western blot of pP65, H3 (histone H3), and GAPDH in nuclei of HUVECs from three independent experiments. c Quantitative analysis of the content of pP65 in nuclei of HUVECs (ratio of pP65 to H3). d Western blot of pP65, P65, and GAPDH in the cytoplasm of HUVECs which is collected from the supernatant after 0.1% Triton treatment in experiment 3. e Representative western blot of pP65, H3, and GAPDH in nuclei of HepG-2 cells. f Quantitative analysis of the content of pP65 in nuclei of HepG-2 cells (ratio of pP65 to H3). The LPS group represents the cells treated with 1 μg/mL LPS for 12 h before western blot. *p < 0.05; **p < 0.01 compared with the commercial kit (a) or with the control (c, f)

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Biological Research

ISSN: 0717-6287

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