Journal: Muscle & Nerve

Article Title: Leucine Supplementation Counteracts the Atrophic Effects of HDAC4 in Rat Skeletal Muscle Submitted to Hindlimb Immobilization

doi: 10.1002/mus.28411

Figure Lengend Snippet: Soleus muscle morphometric analysis after 7 days of hind limb immobilization and leucine supplementation in animals that overexpressed HDAC4. (A) Representative immunofluorescence photomicrographs of soleus muscle, laminin (green) and DAPI (blue, used for nuclei identification) (scale bar 100 μm) and hematoxylin–eosin staining (scale bar 50 μm). The asterisk corresponds to the same fiber in both HE and immunofluorescence technique. (B) Soleus muscle mass, (C) whole muscle CSA, (D) fiber CSA and (E) fiber distribution from EV, pCMVHDAC4, Imm/pCMVHDAC4 and Imm + Leu/pCMVHDAC4 groups. Data are expressed as mean ± SEM. Statistical analysis included the two‐way ANOVA test followed by Tukey's post hoc . * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001 ( n = 5 per group).

Article Snippet: After 30 min, muscles were injected with 50 μL of the HDAC4 expression vector (1.0 μg/μL, pCMVHDAC4, Origene CAT#: RR217458, Rockville, MD, USA) or the empty vector (EV) and an electric pulse generator delivered a burst of pulses (6 pulses of 20 ms with 980 ms pause, 25 V).

Techniques: Immunofluorescence, Staining

Journal: Muscle & Nerve

Article Title: Leucine Supplementation Counteracts the Atrophic Effects of HDAC4 in Rat Skeletal Muscle Submitted to Hindlimb Immobilization

doi: 10.1002/mus.28411

Figure Lengend Snippet: HDAC4 protein expression analysis after 7 days of hind limb immobilization and leucine supplementation in animals that overexpressed HDAC4. (A) HDAC4 and GAPDH western blot showing representative bands. (B) Densitometry analysis by using GAPDH protein level as loading control. Data are expressed as mean ± SEM. Statistical analysis included the two‐way ANOVA test followed by Tukey's post hoc . ** p < 0.01 ( n = 5 per group).

Article Snippet: After 30 min, muscles were injected with 50 μL of the HDAC4 expression vector (1.0 μg/μL, pCMVHDAC4, Origene CAT#: RR217458, Rockville, MD, USA) or the empty vector (EV) and an electric pulse generator delivered a burst of pulses (6 pulses of 20 ms with 980 ms pause, 25 V).

Techniques: Expressing, Western Blot, Control

Journal: Muscle & Nerve

Article Title: Leucine Supplementation Counteracts the Atrophic Effects of HDAC4 in Rat Skeletal Muscle Submitted to Hindlimb Immobilization

doi: 10.1002/mus.28411

Figure Lengend Snippet: Effect of leucine on HDAC4 canonical pathway in animals that overexpressed HDAC4. (A) HDAC4 and (B) Atrogin‐1 mRNA levels were determined by qPCR. (C) Schematic illustrating HDAC4 action in the skeletal muscle fiber nucleus. Cyclophilin A expression was used as housekeeping. Data were expressed as mean ± SEM. Statistical analysis included one‐way ANOVA followed by Tukey's post hoc test. * p < 0.05; ** p < 0.01 ( n = 5 per group).

Article Snippet: After 30 min, muscles were injected with 50 μL of the HDAC4 expression vector (1.0 μg/μL, pCMVHDAC4, Origene CAT#: RR217458, Rockville, MD, USA) or the empty vector (EV) and an electric pulse generator delivered a burst of pulses (6 pulses of 20 ms with 980 ms pause, 25 V).

Techniques: Expressing

Journal: Muscle & Nerve

Article Title: Leucine Supplementation Counteracts the Atrophic Effects of HDAC4 in Rat Skeletal Muscle Submitted to Hindlimb Immobilization

doi: 10.1002/mus.28411

Figure Lengend Snippet: HDAC4 nuclei localization after 7 days of hind limb immobilization and leucine supplementation in animals that overexpressed HDAC4. (A) Representative immunofluorescence photomicrographs of HDAC4 in soleus muscle, HDAC4 (red), DAPI (blue, used for nuclei identification) (scale bar 100 μm). (B) Number of positive HDAC4 nuclei per total nuclei (%). Data were expressed as mean ± SEM. Statistical analysis included two‐way ANOVA followed by Tukey's post hoc test. * p < 0.05; ** p < 0.01 ( n = 5 per group).

Article Snippet: After 30 min, muscles were injected with 50 μL of the HDAC4 expression vector (1.0 μg/μL, pCMVHDAC4, Origene CAT#: RR217458, Rockville, MD, USA) or the empty vector (EV) and an electric pulse generator delivered a burst of pulses (6 pulses of 20 ms with 980 ms pause, 25 V).

Techniques: Immunofluorescence