Journal: PLoS ONE

Article Title: MicroRNAs Overexpressed in Growth-Restricted Rat Skeletal Muscles Regulate the Glucose Transport in Cell Culture Targeting Central TGF-β Factor SMAD4

doi: 10.1371/journal.pone.0034596

Figure Lengend Snippet: PremiR transfections (25 nM) of rat L6 myoblast-myocytes were done and the cell-extracts were subjected to Western Blot analysis for SMAD4, nucleolin and DnaJ-B1 level after 48 hours (I). The triplicate experiments were run in SDS-PAGE/Western Blot to have statistical significant data (T-test) and expressed as relative intensities (%) (I).

Article Snippet: Mammalian skeletal muscle cell-lines (bought from ATCC) of rat origin L6 myoblasts and H9c2 cardiomyocytes (embryonic heart tissue, myoblast morphology) were used in this study.

Techniques: Transfection, Western Blot, SDS Page

Journal: PLoS ONE

Article Title: MicroRNAs Overexpressed in Growth-Restricted Rat Skeletal Muscles Regulate the Glucose Transport in Cell Culture Targeting Central TGF-β Factor SMAD4

doi: 10.1371/journal.pone.0034596

Figure Lengend Snippet: Small inhibitory RNA (25 nM) was used to deplete SMAD4 in L6 cell culture system prior to glucose transport assay using 14 C-2-deoxyglucose as described in . Both skeletal muscle cells (I & II) and cardiomyocytes cells (III) were used for this experiment. The transport assay condition is maintained in the same way as was done for . (IV) The Western Blot analysis in L6 cell-lines to show the specificity of siRNA against SMAD4. Dharmacon-designed SMARTpool siRNAs (4 sets) specific to SMAD4 and non-targeting negative control siRNA were used for this experiment according to the manufacturer's instruction.

Article Snippet: Mammalian skeletal muscle cell-lines (bought from ATCC) of rat origin L6 myoblasts and H9c2 cardiomyocytes (embryonic heart tissue, myoblast morphology) were used in this study.

Techniques: Cell Culture, Transport Assay, Western Blot, Negative Control

Journal: Molecules

Article Title: Anti-Cancer and Anti-Inflammatory Activities of a Short Molecule, PS14 Derived from the Virulent Cellulose Binding Domain of Aphanomyces invadans , on Human Laryngeal Epithelial Cells and an In Vivo Zebrafish Embryo Model

doi: 10.3390/molecules27217333

Figure Lengend Snippet: In vitro toxicity of PS14 tested on rat skeletal myoblast cells (L6 cell line) assessed by MTT assay. The bar shows the toxicity values of control (untreated), PC (positive control) Triton X-100 (0.01%) and PS14-treated experimental sample at five different concentrations. The asterix (*) denotes p < 0.05 level of significance compared to the control. Data were presented as mean ± standard deviation (SD) of three independent experiments.

Article Snippet: Rat skeletal myoblast cells (L6 cell line) and human laryngeal cancer cells (Hep-2) were obtained from the Department of Virology, The King Institute of Preventive Medicine and Research, Guindy, Chennai, India.

Techniques: In Vitro, MTT Assay, Control, Positive Control, Standard Deviation

Journal: Diabetologia

Article Title: Treatment with a β-2-adrenoceptor agonist stimulates glucose uptake in skeletal muscle and improves glucose homeostasis, insulin resistance and hepatic steatosis in mice with diet-induced obesity

doi: 10.1007/s00125-020-05171-y

Figure Lengend Snippet: In vitro and acute in vivo effects of clenbuterol. ( a ) Clenbuterol stimulation of L6 myotubes induced glucose uptake in a dose-dependent manner; n = 5–8; data were analysed by one-way ANOVA with Dunnett’s multiple comparison test. ( b , c ) Stimulation of L6 myotubes stably expressing GLUT4myc with 1 μmol/l clenbuterol induces GLUT4 translocation as quantified in ( c ); n = 3; data were analysed with unpaired two-tailed Student’s t test. Scale bars, 50 μm. ( d ) In vivo glucose uptake in gastrocnemius muscle of chow-fed mice treated with 1 mg/kg clenbuterol for 1 h; n = 5–6; data were analysed with unpaired two-tailed Student’s t test. ( e , f ) Acute effects of clenbuterol on blood glucose ( e ) and plasma insulin ( f ); n = 6 (diet-induced obesity was developed in C57Bl/6N mice maintained at 30°C and on HFD for 7 months; mice were fasted in the morning for 6 h prior to clenbuterol, glucose or saline i.p. injection); data were analysed by two-way ANOVA with Dunnett’s multiple comparison test. In all graphs: * p < 0.05, ** p < 0.01, *** p < 0.001 vs vehicle (in e and f vs vehicle at the same time point)

Article Snippet: L6 rat myoblasts and L6 myoblasts stably expressing GLUT4myc were purchased from KeraFast (ESK201 and ESK202), where they were tested for mycoplasma.

Techniques: In Vitro, In Vivo, Stable Transfection, Expressing, Translocation Assay, Two Tailed Test, Injection

Journal: International Journal of Molecular Sciences

Article Title: Skeletal Muscle Dysfunction in Experimental Pulmonary Hypertension

doi: 10.3390/ijms231810912

Figure Lengend Snippet: ( A ) There is an increased expression of FoxO1 mRNA in the EDL (fast twitch) when this is compared to the soleus (slow twitch) from the control animals. ( B ) A trend for the upregulation of FoxO1 mRNA in soleus of SU/Hx-induced PH is compared to that of the control animals. ( C ) A trend for the upregulation of FoxO1 mRNA in diaphragm of SU/Hx-induced PH is compared to that of the control animals. Expression is normalized to Nup 133 . Values are expressed as the mean ± SEM. Statistical analysis is conducted by a Student’s t -test, ** p < 0.01, ns: not significant. ( D ) There is an increased expression of myogenin and FoxO1 mRNA in the whole cell lysates from differentiated L6 cells (Day 7) when this is compared to the whole cell lysates from undifferentiated cells. Expression is normalized to GAPDH . Values are expressed as the mean ± SEM. Statistical analysis was conducted by a Student’s t -test, ** p < 0.01. ( E ) Immunoblots and quantitative analysis showing that there was a cellular fractionation of the L6 myoblasts and myotube lysates. There was predominantly nuclear FoxO1 localization in myoblasts. There was cytosolic p-FoxO1 localization in myoblasts and myotubes. GAPDH was used as a marker for cytosolic fraction, and H3 was used as a marker for nuclear fraction. Values are expressed as the mean ± SEM. Statistical analysis was conducted by a Student’s t -test, * p < 0.05. ( F ) L6 myotube structure formation (indicated with arrows) after FoxO1 inhibitor treatment (AS1842856, 100 nM, 24 h). Immunoblots showing that there were unchanged levels of type II marker MYH1/2/4/6 and increased levels of type I marker Hexokinase in L6 myotube lysates after FoxO1 inhibitor treatment.

Article Snippet: L6 rat myoblast cells (Kerafast, Boston, MA, USA) were cultured in DMEM that was supplemented with 10% FBS, 100 mg/mL penicillin, and 100 mg/mL streptomycin.

Techniques: Expressing, Western Blot, Cell Fractionation, Marker