Journal: Scientific reports

Article Title: Over-expression of a retinol dehydrogenase (SRP35/DHRS7C) in skeletal muscle activates mTORC2, enhances glucose metabolism and muscle performance.

doi: 10.1038/s41598-017-18844-3

Figure Lengend Snippet: Figure 4. AMPK and Akt phosphorylation levels are increased in muscles from SRP35TG mice. Western blots of total homogenates from EDL and soleus muscles. Phosphorylation of AMPKT172 and AktS473 was determined under the following experimental conditions. (A) and (B) basal conditions (n = 7). (C) and (D) electrical stimulation with a train of tetani (80 Hz, 300 ms duration) delivered at 0.27 Hz for 5 min (n = 6). (E) and (F) stimulation with 100 nM insulin (n = 6). Representatives western blots from the three conditions are show in panels (A, C and E) and the Bar histograms in panels (B, D and F). Fifty µg of protein from total muscle homogenates were loaded per lane, separated on 10% SDS-PAGE and blotted onto nitrocellulose. The immunoreactivity to desmin was used to normalize protein loading. Data are presented as % ± S.D. of WT values (control); *p < 0.05 Student’s t test.

Article Snippet: Subsequently, 50 μg of protein were separated on a 10% SDS PAG, transferred onto nitrocellulose (Amersham), and probed with the following primary antibodies: phospho-AMPK (Thr172), AMPK alpha, phospho-Akt (ser 473), phopho-Akt (thr 308) and Akt total from Cell Signaling and Desmin (used as housekeeping loading control) from Santa Cruz.

Techniques: Phospho-proteomics, Muscles, Western Blot, SDS Page, Control

Journal: Scientific reports

Article Title: Over-expression of a retinol dehydrogenase (SRP35/DHRS7C) in skeletal muscle activates mTORC2, enhances glucose metabolism and muscle performance.

doi: 10.1038/s41598-017-18844-3

Figure Lengend Snippet: Figure 5. atRA activates AktS473 phosphorylation in WT EDL and soleus muscles. Fifty µg of protein from total muscle homogenates were loaded per lane, separated on 10% SDS-PAGE and blotted onto nitrocellulose. AMPKT172, AktS473 and AktT308 phosphorylation levels were measured by western blotting on total homogenates from EDL and soleus with specific anti-phospho Ab; P/T = Phosphorylated protein/Total protein. The immunoreactivity to desmin was used to normalize gel loading. Data are presented as % ( ± S.D.) of control (empty bar); * p < 0.05 Student’s t test. (A) and (C) Skeletal muscles isolated from WT mice were incubated with 10 μM all-trans-retinoic acid (atRA, bars; n = 6). Left side: representative western blots at 30 min of incubation; right side: bar histogram plots. Control values (empty bar) for normalization were obtained by incubating the contralateral muscle in the presence of the vehicle solution (DMSO). (B) and (D) EDL and soleus muscles incubated with 100 nM insulin plus or minus 10 μM atRA. Left side: representative western blots at 30 min of incubation; right side: bar histogram plots (n = 6 *p < 0.05 Student’s t test). Control values (empty bar) for normalization were obtained by performing the experiments in the presence of 100 nM insulin plus vehicle (DMSO). (E) and (F) EDL muscles were incubated for 30 min in presence of 100 nM insulin, RARα, RARβ and RARγ inhibitors plus or minus 10 μM atRA. Control values (empty bar) were obtained in the presence of 100 nM insulin, plus the RAR inhibitor. Data are presented as mean ± S.D., n = 6, *p < 0.05 Student’s t test.

Article Snippet: Subsequently, 50 μg of protein were separated on a 10% SDS PAG, transferred onto nitrocellulose (Amersham), and probed with the following primary antibodies: phospho-AMPK (Thr172), AMPK alpha, phospho-Akt (ser 473), phopho-Akt (thr 308) and Akt total from Cell Signaling and Desmin (used as housekeeping loading control) from Santa Cruz.

Techniques: Phospho-proteomics, Muscles, SDS Page, Western Blot, Control, Isolation, Incubation

Journal: Scientific reports

Article Title: Over-expression of a retinol dehydrogenase (SRP35/DHRS7C) in skeletal muscle activates mTORC2, enhances glucose metabolism and muscle performance.

doi: 10.1038/s41598-017-18844-3

Figure Lengend Snippet: Figure 6. SRP-35 and atRA activation of AktS473 is controlled by mTORC2 and the PI3K signaling pathway. (A) and (B) Western blot images and bar histograms showing the phosphorylation of Akt and AMPK in EDL and soleus muscles from WT (n = 5) and RimKO (n = 5) mice incubated with or without 10 μM atRA in presence of 100 nM insulin. Fifty µg of total homogenate protein were loaded per lane, separated on 10% SDS/PAGE and blotted onto nitrocellulose. P/T = Phosphorylated protein/Total protein. Anti-desmin immunoreactivity was used as loading control (bars represent the mean ± S.D., *p < 0.05 Student’s t test). (C) PIP3 levels from EDL muscles isolated from WT and SRP35TG mice fed a standard chow diet (Top panel) and on a low vitamin A diet (LVA, 4 I.U/Kg, Bottom panel). Each symbol represents PIP3 values from a single mouse; the median value of the data is shown in the box-plot; n.d. = non detectable (muscles from 7 mice) *p < 0.05 Mann-Whitney test. (D) PIP3 levels from soleus muscles isolated from WT and SRP35TG mice fed a standard chow diet (Top panel) and on a low vitamin A diet (LVA, 4 I.U/Kg, Bottom panel). Each symbol represents PIP3 values from a single mouse; the median value of the data is shown in the box-plot; *p < 0.05 Mann -Whitney test. (E) and (F) EDL

Article Snippet: Subsequently, 50 μg of protein were separated on a 10% SDS PAG, transferred onto nitrocellulose (Amersham), and probed with the following primary antibodies: phospho-AMPK (Thr172), AMPK alpha, phospho-Akt (ser 473), phopho-Akt (thr 308) and Akt total from Cell Signaling and Desmin (used as housekeeping loading control) from Santa Cruz.

Techniques: Activation Assay, Western Blot, Phospho-proteomics, Muscles, Incubation, SDS Page, Control, Isolation, MANN-WHITNEY

Journal: Cytoskeleton (Hoboken, N.J.)

Article Title: Disassembly of Myofibrils and Potential Imbalanced Forces on Z-Discs in Cultured Adult Cardiomyocytes

doi: 10.1002/cm.21298

Figure Lengend Snippet: (A) The patterns of desmin in the different shapes of cardiomyocytes. The striated pattern was still visible at the middle region (red arrows), but disappeared at the ends of a peanut-shape cell (yellow arrows). (B) Striated myofibril structure is clearly shown in a LRS cardiomyocyte, and the patterns of desmin and α-actinin are co-localized. (C) The striated myofibril patterns disappeared at the cell ends (yellow arrows) and the patterns of desmin and α-actinin became mismatched in peanut-shape cells. At the middle region of the cell (red arrows), the striated patterns of desmin and α-actinin are still visible and co-localized. Scale bar: 10 μm.

Article Snippet: After permeabilizing the cells using Triton X-100 and blocking the active sites in blocking solution (0.2% Triton X-100, 5% normal serum and 1% bovine serum albumin in phosphate buffered saline) for 30 min at room temperature, primary antibodies, including anti-α-actinin (Abcam), anti-connexin-43 (Santa Cruz), anti-MyHC (Abcam), anti-plakoglobin, and anti-desmin (Novus Biologicals) were applied.

Techniques:

Journal: Cytoskeleton (Hoboken, N.J.)

Article Title: Disassembly of Myofibrils and Potential Imbalanced Forces on Z-Discs in Cultured Adult Cardiomyocytes

doi: 10.1002/cm.21298

Figure Lengend Snippet: (A) Representative images of cardiomyocytes cultured in medium with or without 25 μM BLB at different time points (Day 0–4). Scale bar: 100 μm. (B) The percentage of LRS cardiomyocytes. *, P < 0.05 versus Control. (C-D) BLB maintained the distribution of α-actinin and preserved desmin and myofibril structure. Scale bar: 50 μm. CTRL: control, BLB: Blebbistatin.

Article Snippet: After permeabilizing the cells using Triton X-100 and blocking the active sites in blocking solution (0.2% Triton X-100, 5% normal serum and 1% bovine serum albumin in phosphate buffered saline) for 30 min at room temperature, primary antibodies, including anti-α-actinin (Abcam), anti-connexin-43 (Santa Cruz), anti-MyHC (Abcam), anti-plakoglobin, and anti-desmin (Novus Biologicals) were applied.

Techniques: Cell Culture

Journal: Cytoskeleton (Hoboken, N.J.)

Article Title: Disassembly of Myofibrils and Potential Imbalanced Forces on Z-Discs in Cultured Adult Cardiomyocytes

doi: 10.1002/cm.21298

Figure Lengend Snippet: (A-C) Freshly isolated cardiomyocytes connected through intercalated discs. (A) and (B) Cardiomyocytes with well-preserved intercalated discs. (C) Cardiomyocytes with damaged intercalated discs. The local movement in the cardiomyocytes was analyzed using contractile image analysis. (A-B) Contraction wave (white rectangle in A) in both cardiomyocytes was well-synchronized at the intercalated discs (white arrows). Similar local displacement fields at each side of well-preserved intercalated discs (B). Completely different local displacement fields at each side of damaged intercalated discs (C). (D-E) The cardiomyocytes (Day 1) were double stained with α-actinin (green) and connexin 43 (C×43) (red) (D), desmin (green) and plakoglobin (red) (E), respectively. Blue: nucleus. Yellow arrows indicate disassembly of myofibrils at the cell ends. Scale bar: 50 μm.

Article Snippet: After permeabilizing the cells using Triton X-100 and blocking the active sites in blocking solution (0.2% Triton X-100, 5% normal serum and 1% bovine serum albumin in phosphate buffered saline) for 30 min at room temperature, primary antibodies, including anti-α-actinin (Abcam), anti-connexin-43 (Santa Cruz), anti-MyHC (Abcam), anti-plakoglobin, and anti-desmin (Novus Biologicals) were applied.

Techniques: Isolation, Staining

Journal: Indian Journal of Pathology and Microbiology

Article Title: Clinical characteristics and gene mutation analysis of clear cell tumor of the lung

doi: 10.4103/ijpm.ijpm_65_19

Figure Lengend Snippet: Figure 3: A1‑3. IHC results revealed positive results in CK (pan) (‑), HMB45 (+), SMA (+) supporting a diagnosis of clear cell tumor of the lung. B1‑3: IHC results revealed positive results in Calponin (+), Melan‑A (+), Melanoma (+), supporting a diagnosis of clear cell tumor of the lung. C1‑3: IHC results revealed positive results in Melanoma (+) and negative results in CK‑pan and Desmin, indicating a diagnosis of clear cell tumor of the lung. D1‑3: IHC results revealed positive results in Melanoma (+), S‑100 (+), SMA (+)

Article Snippet: The antibodies used were as follows: Calponin (EP63, ZSBIO, Beijing, China), Melanoma (HMB‐45, MAXIM, Fuzhou, China), S100 (15E2E2 + 4C.9, ZSBIO, Beijing, China), CK‐pan (AE1, MAXIM, Fuzhou, China), Desmin (RD301, MAXIM, Fuzhou, China), SMA (SCB3, ZSBIO, Beijing, China), Melan‐A (SPM540, ZSBIO, Beijing, China).

Techniques: Biomarker Discovery