Research Paper Volume 3, Issue 10 pp 955—967

Figure 4. Co-staining of total protein and histone macro H2A. Cell we stained with NanoOrange (NO) and subsequently processed for immunofluorescent detection of histone macro H2A (mH2A) as indicated in Methods. (A) Representative images acquired with cultures of HDF. The panels are arranged and labeled as indicated in Figure 2A. (B) Histograms of mH2A staining for early passage and senescent HDF cells. The data are displayed as indicated in Figure 2B. (C) Mean values of staining intensity for DNA, total protein and mH2A in early passage and senescent HDF cells. The data are displayed as indicated in Figure 2C. The increases of both protein and mH2A signals are statistically significant (* p < 0.001, ** p < 0.001). The signal for DNA did not change significantly between early passage and senescence (p = 0.34). (D) Representative images acquired with cultures of MTF. The panels are arranged and labeled as indicated in (A) above. (E) Histograms of mH2A staining for early passage and senescent MTF cells. The data are displayed as indicated in (B) above. (F) Mean values of staining intensity for DNA, total protein and mH2A in early passage and senescent MTF cells. The data are displayed as indicated in (C) above. As with the HDF, the increases in both protein and mH2A signals were statistically significant (* p < 0.001, ** p < 0.001), whereas the DNA signal did not change significantly (* p = 0.29).