Histone H3 Antibody - #BF9211

Fig. 2 The activation of SIRT1 by RES improved DOX-induced inflammation in mice hearts and H9c2 cells. (A) Cardiac inflammatory response was detected by IHC staining for tumor necrosis factor-α (TNF-α, brown considered positive staining) followed by a quantitative analysis of the positive stains (n = 6). (B) Relative myocardial interleukin-1β (Il1b) mRNA level was measured by RT-qPCR (n = 5–6). (C, D) H9c2 cells were transfected with NC-shRNA or Sirt1-shRNA for 24 h, and then treated with RES (20 μM) or DOX (1 μM) or both for 24 h. Relative Tnfa and Il1b mRNA levels were measured by RT-qPCR. Three independent experiments were performed. (E) Western blot analysis and densitometric quantification of nuclear nuclear factor κB p65 (NF-κB p65) expression in cardiac tissues of each group (n = 6). Histone H3 as an internal control. (F) Nuclear accumulation of NF-κB p65 (indicated by white arrows) determined by immunofluorescent staining with anti-NF-κB p65 antibody (red) in cardiac tissues. (G) Western blot analysis and densitometric quantification of nuclear NF-κB p65 expression in H9c2 cells of different groups. Three independent experiments were performed. Histone H3 as an internal control. Data were presented as mean ± SD. *P < 0.05, ns indicates no significance. (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)

Figure 4 The activation of NRF2 was elevated after RES and FGF1 co-treating in DOX-treated mice. (A–C) The expression of NRF2, HO-1 and NQO1 was detected by immunofluorescent staining in cardiac tissues, respectively. (D–G) The levels of relative protein expression were examined by Western blot assay with densitometric quantification (Ctrl: n = 4; other groups: n = 6). (H–J) Representative images of DHE (red) and DCFH-DA (green) staining and quantification of fluorescence intensity in H9c2 cells (n = 3). GAPDH or Histone H3 was used as loading controls for all Western blot assays. Data are expressed as means ± SD.

FIGURE 1 Foam cells are characterized by autophagy deficiency and lysosomal dysfunction. A-C, Protein levels of LC3I/II and p62 were determined by Western blotting. Representative Western blotting A, and quantitative analysis B and C, displayed a reduced protein level of LC3II and increased protein level of p62. D, The mRNA expression levels of autophagic genes were determined by RT-PCR. E, Foam cells displayed reduced autolysosomes as shown by the reduced red puncta of mRFP-GFP-LC3. F-H, Measurement of lysosomal biogenesis and function. F, LysoTracker staining for lysosomal biogenesis. G, Protein level of CTSD for lysosomal degradation capability. H, LysoSensor staining for pH. *P < .05; **P < .01. Results are means ± SD of three independent experiments. The value represents fold of vehicle