Journal: Cell metabolism

Article Title: Elevated stearoyl-CoA desaturase-1 expression in skeletal muscle contributes to abnormal fatty acid partitioning in obese humans

doi: 10.1016/j.cmet.2005.09.002

Figure Lengend Snippet: Preconfluent myoblasts were transiently transfected with pcDNA3.0-hSCD1 or the empty parent vector. After 7 days in DFM, cells were harvested to obtain total RNA. Overexpression of SCD1 was confirmed in differentiated myotubes by (A) semiquantitative PCR of mRNA and (B) Western blot analysis of protein. β-actin mRNA and GAPDH protein were used as loading controls. Metabolic assays were performed following a 3 hr incubation with 100 μM [14C]oleate and 0.25% BSA. (C) Fatty acid oxidation was determined by measuring 14C-label incorporation into CO2. (D) Glycerolipid synthesis was determined by measuring 14C-label incorporation into triacylglycerol (TAG) and phospholipid (PL). All assays were performed in triplicate and values represent the mean ± SEM from four separate experiments. Differences between groups were analyzed by Student’s t test, *p < 0.05, **p < 0.01. (E) ACCβ mRNA levels were quantified by RTQ-PCR and are expressed as relative units in cells transfected with SCD1 versus empty vector controls. Cyclophilin B was measured as an endogenous control. Data are representative of five experiments and differences between groups were analyzed by Student’s t test, *p < 0.05. (F) Total and phosphorylated AMPK and ACCβ were analyzed by immunoblotting with antibodies against the AMPKα 1 and 2 catalytic subunits, ACCβ, AMPK-thr172, and ACCβ-ser79. GAPDH protein expression was not different between groups.

Article Snippet: Blots were probed with primary antibodies against hSCD1 (4 μg/mL, Alpha Diagnostics International, San Anto-nio, TX) and GAPDH (1:100 dilution, Santa Cruz Biotechnology, Santa Cruz, CA) followed by anti-rabbit and anti-goat secondary antibodies respectively (1:8000 and 1:10000 dilutions, respectively; Santa Cruz Biotechnology, Santa Cruz, CA).

Techniques: Transfection, Plasmid Preparation, Over Expression, Western Blot, Incubation, Control, Expressing

Journal: Pain

Article Title: Peripheral oxytocin restores light touch and nociceptor sensory afferents towards normal after nerve injury

doi: 10.1097/j.pain.0000000000001495

Figure Lengend Snippet: A. Representative vasopressin 1A receptor immunoreactivity in sections from L4 dorsal root ganglia (DRGs) visualized by fluorescence microscopy (vasopressin 1a receptors in green and NeuN in red). Calibration bar: 50 μm.

Article Snippet: The anti-vasopressin 1A receptor antibody (rabbit, 1:100, #AVP1A11-P, Alpha Diagnostics, San Antonio, TX, USA) showed selective immunostaining in brain regions of vasopressin 1A receptor location in the rat brain and consistent immunostaining in a subset of dorsal root ganglion neurons and was used for tissues from animals in the experiment.

Techniques: Fluorescence, Microscopy

Journal: Journal of Cellular and Molecular Medicine

Article Title: Byakangelicin protects against carbon tetrachloride–induced liver injury and fibrosis in mice

doi: 10.1111/jcmm.15493

Figure Lengend Snippet: Byakangelicin treatment reduces carbon tetrachloride‐induced liver fibrosis. A, Animal liver fibrosis modelling plan. B, Detection of hydroxyproline in liver tissue. C, Classical tissue staining in liver fibrosis: Sirius red, Masson and α‐SMA staining; D, three stained quantitative areas. E, Protein content of α‐SMA and COL‐1 in liver tissue; F, corresponding grey analysis. G, Real‐time PCR analyses of α‐SMA and COL‐1 in liver tissues. For the statistics of each panel in this figure, * P < .05 vs model, ** P < .01 vs model, *** P < .001 vs model, n = 3

Article Snippet: Primary antibodies including anti‐α‐smooth muscle actin (α‐SMA), anti‐collagen I (COL‐1), anti‐GAPDH and anti‐p‐Stat3, Stat3, β‐tubulin, cyclin D1, P53, p‐ASK‐1, ASK‐1, IL‐1β, NF‐κB and Nrf‐2 were acquired from Affinity Biosciences; anti‐p‐Smad3, Smad3, p‐PDGFR, PDGFR, p‐ERK, ERK, p‐AKT, AKT, PARP, cleaved caspase‐3 and caspase‐3 were received from (Cell Signaling Technology); anti‐p‐JNK, JNK from Signalway Antibody LLC; and anti‐4‐HNE from Abcam.

Techniques: Staining, Real-time Polymerase Chain Reaction

Journal: Journal of Cellular and Molecular Medicine

Article Title: Byakangelicin protects against carbon tetrachloride–induced liver injury and fibrosis in mice

doi: 10.1111/jcmm.15493

Figure Lengend Snippet: Byakangelicin inhibits the TGF/Smad3 pathway in TGF‐β‐induced activation of hepatic stellate cell. A, Detection of byakangelicin's cytotoxicity in liver stellate cell line LX2 using MTT. B, Real‐time PCR analyses of α‐SMA and COL‐1 in LX2 cells. C and D, Western blot analyses of α‐SMA, COL‐1, P‐Smad3, Smad3 and GAPDH protein expression in LX2 cells with densitometry. E and F, Immunofluorescence using antibody against α‐SMA and Smad3. For the statistics of each panel in this figure, * P < .05, ** P < .01, *** P < .001, n = 3

Article Snippet: Primary antibodies including anti‐α‐smooth muscle actin (α‐SMA), anti‐collagen I (COL‐1), anti‐GAPDH and anti‐p‐Stat3, Stat3, β‐tubulin, cyclin D1, P53, p‐ASK‐1, ASK‐1, IL‐1β, NF‐κB and Nrf‐2 were acquired from Affinity Biosciences; anti‐p‐Smad3, Smad3, p‐PDGFR, PDGFR, p‐ERK, ERK, p‐AKT, AKT, PARP, cleaved caspase‐3 and caspase‐3 were received from (Cell Signaling Technology); anti‐p‐JNK, JNK from Signalway Antibody LLC; and anti‐4‐HNE from Abcam.

Techniques: Activation Assay, Real-time Polymerase Chain Reaction, Western Blot, Expressing, Immunofluorescence

Journal: Journal of Cellular and Molecular Medicine

Article Title: Byakangelicin protects against carbon tetrachloride–induced liver injury and fibrosis in mice

doi: 10.1111/jcmm.15493

Figure Lengend Snippet: Byakangelicin inhibits the proliferation and activation of PDGF‐induced hepatic stellate cell by inhibiting PDGFR/ERK, PDGFR/AKT, and PDGFR/Stat3. A and B, Western blot analyses of P‐PDGFR, PDGFR, P‐ERK, ERK, P‐AKT, AKT, P‐Stat3, Stat3, α‐SMA and cyclin D1 protein expression with densitometry. C and D, Immunofluorescence by using antibody against cyclin D1 and PDGFR. For the statistics of each panel in this figure, * P < .05, ** P < .01, *** P < .001, n = 3

Article Snippet: Primary antibodies including anti‐α‐smooth muscle actin (α‐SMA), anti‐collagen I (COL‐1), anti‐GAPDH and anti‐p‐Stat3, Stat3, β‐tubulin, cyclin D1, P53, p‐ASK‐1, ASK‐1, IL‐1β, NF‐κB and Nrf‐2 were acquired from Affinity Biosciences; anti‐p‐Smad3, Smad3, p‐PDGFR, PDGFR, p‐ERK, ERK, p‐AKT, AKT, PARP, cleaved caspase‐3 and caspase‐3 were received from (Cell Signaling Technology); anti‐p‐JNK, JNK from Signalway Antibody LLC; and anti‐4‐HNE from Abcam.

Techniques: Activation Assay, Western Blot, Expressing, Immunofluorescence