rabbit anti-rat histone 3 antibody  (Santa Cruz Biotechnology)

Journal: International Journal of Molecular Medicine

Article Title: Biphasic activation of nuclear factor-κB and expression of p65 and c-Rel following traumatic neuronal injury

doi: 10.3892/ijmm.2018.3567

Figure Lengend Snippet: Western blotting was performed to detect the nuclear protein levels of p65 and c-Rel in cultured neurons. (A) Representative western blots show the p65 and c-Rel protein levels of the neurons in all groups. Histone 3 was used as the loading control. Nuclear protein levels of (B) p65 and (C) c-Rel in the cultured neurons were markedly increased in the transection groups compared with those in the control group, showing different activated phases. The p65 protein levels exhibited two peaks at 1 and 24 h, while c-Rel levels were significantly elevated during the later stage post-injury. Data are presented as mean ± standard error of the mean (n=6 per group). * P<0.05, ** P<0.01 and *** P<0.001 vs. control group.

Article Snippet: The primary antibodies were rabbit anti-rat p65 (sc-372) and c-Rel (sc-272) (1:200 dilution), and rabbit anti-rat histone 3 antibody (sc-8654; 1:200 dilution; each from Santa Cruz Biotechnology, Inc., Dallas, TX, USA) was used as a loading control.

Techniques: Western Blot, Cell Culture

Journal: American Journal of Physiology - Regulatory, Integrative and Comparative Physiology

Article Title: Sepsis and glucocorticoids upregulate p300 and downregulate HDAC6 expression and activity in skeletal muscle

doi: 10.1152/ajpregu.00858.2009

Figure Lengend Snippet: Histone deacetylases (HDAC)3 and -6 mRNA levels in skeletal muscle during sepsis in rats. HDAC3 (A) and HDAC6 (B) mRNA levels were determined by real-time PCR in EDL muscles at different time points after sham operation or induction of sepsis by CLP in rats. Results are means ± SE with n = 8 in each group. Statistical analysis was performed using ANOVA.

Article Snippet: The membranes were blocked with 5% nonfat milk in TTBS buffer (50 mM Tris·HCl, 150 mM NaCl, and 1% Tween-20, pH 7.4) and incubated with the following primary antibodies and the appropriate secondary antibodies: a rabbit polyclonal anti-rat HDAC3 antibody (1:1,000, Cell Signaling Technology, Danver, MA); a rabbit polyclonal anti-rat HDAC6 antibody (1:1,000, Santa Cruz Biotechnology, Santa Cruz, CA); a rabbit polyclonal anti-rat SIRT1 antibody (1:500, Abcam, Cambridge, MA); a rabbit polyclonal anti-mouse MAFbx/atrogin-1 antibody (1:1,000; kindly supplied by Dr. Stewart Lecker, Harvard Medical School); and a rabbit polyclonal anti-mouse MuRF1 antibody (kindly supplied by Regeneron Pharmaceuticals to Dr. Alfred Goldberg, Harvard Medical School, and used in the present experiments after permission from Regeneron Pharmaceuticals).

Techniques: Real-time Polymerase Chain Reaction, Muscles

Journal: American Journal of Physiology - Regulatory, Integrative and Comparative Physiology

Article Title: Sepsis and glucocorticoids upregulate p300 and downregulate HDAC6 expression and activity in skeletal muscle

doi: 10.1152/ajpregu.00858.2009

Figure Lengend Snippet: Sepsis in rats reduces protein levels of HDAC6 and SIRT1 (a class III HDAC) in skeletal muscle. Protein levels of HDAC3 (A), HDAC6 (B), and SIRT1 (C) were determined by Western blot analysis of nuclear proteins. OCT1 or lamin A/C levels were determined for loading control. Representative blots are shown. Bars represent densitometric quantifications of 7 or 8 blots. Results are means ± SE. *P < 0.05 vs. sham determined by Student's t-test.

Article Snippet: The membranes were blocked with 5% nonfat milk in TTBS buffer (50 mM Tris·HCl, 150 mM NaCl, and 1% Tween-20, pH 7.4) and incubated with the following primary antibodies and the appropriate secondary antibodies: a rabbit polyclonal anti-rat HDAC3 antibody (1:1,000, Cell Signaling Technology, Danver, MA); a rabbit polyclonal anti-rat HDAC6 antibody (1:1,000, Santa Cruz Biotechnology, Santa Cruz, CA); a rabbit polyclonal anti-rat SIRT1 antibody (1:500, Abcam, Cambridge, MA); a rabbit polyclonal anti-mouse MAFbx/atrogin-1 antibody (1:1,000; kindly supplied by Dr. Stewart Lecker, Harvard Medical School); and a rabbit polyclonal anti-mouse MuRF1 antibody (kindly supplied by Regeneron Pharmaceuticals to Dr. Alfred Goldberg, Harvard Medical School, and used in the present experiments after permission from Regeneron Pharmaceuticals).

Techniques: Western Blot, Control

Journal: American Journal of Physiology - Regulatory, Integrative and Comparative Physiology

Article Title: Sepsis and glucocorticoids upregulate p300 and downregulate HDAC6 expression and activity in skeletal muscle

doi: 10.1152/ajpregu.00858.2009

Figure Lengend Snippet: Treatment of rats with dexamethasone reduces the expression of HDAC3 and -6 and inhibits HDAC activity in skeletal muscle. HDAC3 (A) and HDAC6 (B) mRNA levels and HDAC activity (C) were determined in EDL muscles 8 and 16 h after treatment of rats with 10 mg/kg of dexamethasone or vehicle (control). Results are means ± SE with n = 8 in each group. *P < 0.05 vs. control at the corresponding time point by ANOVA.

Article Snippet: The membranes were blocked with 5% nonfat milk in TTBS buffer (50 mM Tris·HCl, 150 mM NaCl, and 1% Tween-20, pH 7.4) and incubated with the following primary antibodies and the appropriate secondary antibodies: a rabbit polyclonal anti-rat HDAC3 antibody (1:1,000, Cell Signaling Technology, Danver, MA); a rabbit polyclonal anti-rat HDAC6 antibody (1:1,000, Santa Cruz Biotechnology, Santa Cruz, CA); a rabbit polyclonal anti-rat SIRT1 antibody (1:500, Abcam, Cambridge, MA); a rabbit polyclonal anti-mouse MAFbx/atrogin-1 antibody (1:1,000; kindly supplied by Dr. Stewart Lecker, Harvard Medical School); and a rabbit polyclonal anti-mouse MuRF1 antibody (kindly supplied by Regeneron Pharmaceuticals to Dr. Alfred Goldberg, Harvard Medical School, and used in the present experiments after permission from Regeneron Pharmaceuticals).

Techniques: Expressing, Activity Assay, Muscles, Control

Journal: American Journal of Physiology - Regulatory, Integrative and Comparative Physiology

Article Title: Sepsis and glucocorticoids upregulate p300 and downregulate HDAC6 expression and activity in skeletal muscle

doi: 10.1152/ajpregu.00858.2009

Figure Lengend Snippet: Treatment of rats with the glucocorticoid receptor antagonist RU38486 prevents sepsis-induced changes in p300 and HDAC6 expression in skeletal muscle. A: p300 mRNA levels were determined by real-time PCR in EDL muscles 16 h after sham operation or CLP. Rats were treated with 10 mg/kg of RU38486 or vehicle (control) by intraperitoneal injection 2 h before sham operation or CLP. HDAC3 (B) and HDAC6 (C) mRNA levels were determined in EDL muscles 8 h after sham operation or CLP. Rats were treated with 10 mg/kg of RU38486 or vehicle (control) by intraperitoneal injection 2 h before sham operation or CLP. Results are means ± SE with n = 8 in each group. *P < 0.05 vs. control by ANOVA.

Article Snippet: The membranes were blocked with 5% nonfat milk in TTBS buffer (50 mM Tris·HCl, 150 mM NaCl, and 1% Tween-20, pH 7.4) and incubated with the following primary antibodies and the appropriate secondary antibodies: a rabbit polyclonal anti-rat HDAC3 antibody (1:1,000, Cell Signaling Technology, Danver, MA); a rabbit polyclonal anti-rat HDAC6 antibody (1:1,000, Santa Cruz Biotechnology, Santa Cruz, CA); a rabbit polyclonal anti-rat SIRT1 antibody (1:500, Abcam, Cambridge, MA); a rabbit polyclonal anti-mouse MAFbx/atrogin-1 antibody (1:1,000; kindly supplied by Dr. Stewart Lecker, Harvard Medical School); and a rabbit polyclonal anti-mouse MuRF1 antibody (kindly supplied by Regeneron Pharmaceuticals to Dr. Alfred Goldberg, Harvard Medical School, and used in the present experiments after permission from Regeneron Pharmaceuticals).

Techniques: Expressing, Real-time Polymerase Chain Reaction, Muscles, Control, Injection

Journal: American Journal of Physiology - Regulatory, Integrative and Comparative Physiology

Article Title: Sepsis and glucocorticoids upregulate p300 and downregulate HDAC6 expression and activity in skeletal muscle

doi: 10.1152/ajpregu.00858.2009

Figure Lengend Snippet: Sepsis-induced changes in soleus muscles of p300, HDAC3, HDAC6, atrogin-1, and MuRF1 mRNA levels. The mRNA levels were determined by real-time PCR at different time points after sham operation or CLP. Results are means ± SE with n = 6–8 in each group. *P < 0.05 vs. sham by ANOVA.

Article Snippet: The membranes were blocked with 5% nonfat milk in TTBS buffer (50 mM Tris·HCl, 150 mM NaCl, and 1% Tween-20, pH 7.4) and incubated with the following primary antibodies and the appropriate secondary antibodies: a rabbit polyclonal anti-rat HDAC3 antibody (1:1,000, Cell Signaling Technology, Danver, MA); a rabbit polyclonal anti-rat HDAC6 antibody (1:1,000, Santa Cruz Biotechnology, Santa Cruz, CA); a rabbit polyclonal anti-rat SIRT1 antibody (1:500, Abcam, Cambridge, MA); a rabbit polyclonal anti-mouse MAFbx/atrogin-1 antibody (1:1,000; kindly supplied by Dr. Stewart Lecker, Harvard Medical School); and a rabbit polyclonal anti-mouse MuRF1 antibody (kindly supplied by Regeneron Pharmaceuticals to Dr. Alfred Goldberg, Harvard Medical School, and used in the present experiments after permission from Regeneron Pharmaceuticals).

Techniques: Muscles, Real-time Polymerase Chain Reaction