Journal: PLoS ONE

Article Title: A Small Insulinomimetic Molecule Also Improves Insulin Sensitivity in Diabetic Mice

doi: 10.1371/journal.pone.0169809

Figure Lengend Snippet: (A) dmp fails to activate EGFR. L6 myotubes or 3T3L1 adipocytes were treated with or without 250 nM dmp for 4h. The cell lysates were analyzed by immunoblotting with anti-pEGFR and anti-EGFR antibodies. (B,C) dmp can induce IR phosphorylation in a dose dependent manner. L6 myotubes were treated with insulin (20–120 n m ) or dmp (50–300 n m ) for 4h and IR phosphorylation was monitored by ELISA (B) or immunoblotting with anti-pIR and anti-IR antibodies (C). (D) IR kinase activity was determined in L6 myotubes which were incubated with varied concentrations of insulin or dmp. (E) dmp stimulates IR and its downstream kinases phosphorylation. L6 myotubes or 3T3L1 adipocytes were treated with or without Insulin (100 nM) or dmp (250 nM) for 4h and the IR phosphorylation and its downstream signalling were monitored by immunoblotting. (F) L6 myotubes transfected with GFP-GLUT4 chimeric gene were incubated with insulin (100 nM) or dmp (250nM) for 4h. Cells on the cover slips were fixed in paraformaldehyde and observed under florescent microscope for GFP-GLUT4 translocation. (G) dmp like insulin promotes glucose uptake. L6 myotubes or skeletal muscle cells from soleus muscle of neonatal mice (2-3days) were incubated with 100 n m insulin or 250 n m dmp for 25 min. [ 14 C] 2-DOG was then added, and the cells were further incubated for 5 min. [ 14 C] 2-DOG uptake was measured by scintillation counting. * P <0.05 versus Con; ** P <0.01 versus Con. (H) dmp augments fatty acid uptake. Primary culture adipocytes or 3T3L1 adipocytes were treated with 100 n m insulin or 250 n m dmp for 4h followed by incubation with [ 3 H] Palmitate for 15 min. [ 3 H] Palmitate uptake was measured in a liquid scintillation counter. * P <0.05 versus Con; ** P <0.01 versus Con. (I) L6 myotubes were tranfected with IR siRNA(IR KD ) followed by estimation of IR gene and protein levels by qPCR (left) and immunoblotting (right) respectively. * P <0.05 versus Con. (J) IR KD L6 myotubes were incubated with dmp for 4h and [ 14 C] 2-DOG uptake was measured according to the above description. ** P <0.01 versus Con. All values are represented as mean ± s.e.m. (n = 5).

Article Snippet: L6 skeletal muscle cell line and mouse pre-adipocyte 3T3L1 were procured from the National Centre for Cell Science, Pune, India.

Techniques: Western Blot, Phospho-proteomics, Enzyme-linked Immunosorbent Assay, Activity Assay, Incubation, Transfection, Microscopy, Translocation Assay

Journal: PLoS ONE

Article Title: A Small Insulinomimetic Molecule Also Improves Insulin Sensitivity in Diabetic Mice

doi: 10.1371/journal.pone.0169809

Figure Lengend Snippet: Rats were fed with dmp (5 mg kg bw -1 ) for 14 days. (A) Hematological, (B) Biochemical analysis, (C) Liver and kidney histopathology were performed on 0 and 14 day to test toxic effect of dmp. (D) L6 myotubes and 3T3L1 adipocytes were incubated with 300 nM of dmp for 4 h and cell viability was assessed by MTT assay. (E) After incubation of 3T3L1 adipocytes with or without 300 nM dmp for 4 h, release of TNFα and IL6 in to the medium was estimated through ELISA. (F) ATP production was measured in L6 myotubes incubated in the absence or presence of 300nM dmp for 4 h. All values are represented as mean ± s.e.m. (n = 5)

Article Snippet: L6 skeletal muscle cell line and mouse pre-adipocyte 3T3L1 were procured from the National Centre for Cell Science, Pune, India.

Techniques: Histopathology, Incubation, MTT Assay, Enzyme-linked Immunosorbent Assay

Journal: BMC Complementary and Alternative Medicine

Article Title: MD-1, a poly herbal formulation indicated in diabetes mellitus ameliorates glucose uptake and inhibits adipogenesis – an in vitro study

doi: 10.1186/s12906-018-2177-x

Figure Lengend Snippet: Cytotoxicity assay in L6 myotubes & 3T3L1 adipocytes. Effect of HAEF on cell viability of ( a ) L6 myotubes after 24 h exposure ( b ) 3T3L1 adipocytes after 7 days exposure. All the data were expressed as means ± S.E.M. of three independent experiments

Article Snippet: L6 myocytes and 3T3L1 preadipocytes were procured from the National Centre for Cell Science, Pune and maintained in Dulbeccos Modified Eagle Medium (DMEM) containing high glucose with 10% fetal bovine serum (FBS) and supplemented with penicillin (5 units/mL) and streptomycin (5 μg/mL) in 5% CO 2 incubator at 37 °C.

Techniques: Cytotoxicity Assay

Journal: BMC Complementary and Alternative Medicine

Article Title: MD-1, a poly herbal formulation indicated in diabetes mellitus ameliorates glucose uptake and inhibits adipogenesis – an in vitro study

doi: 10.1186/s12906-018-2177-x

Figure Lengend Snippet: Effect of HAEF on Glucose uptake in L6 myotubes & 3T3L1 adipocytes. Effect of HAEF on glucose uptake on ( a ) L6 myotubes & ( b ) 3T3L1 adipocytes. The cells were treated with different concentrations of HAEF and pioglitazone (PGZ) for 24 h in the presence and absence of insulin. All the data were expressed as means ± S.E.M. of three independent experiments. * P < 0.05 as compared with respective (−) insulin group

Article Snippet: L6 myocytes and 3T3L1 preadipocytes were procured from the National Centre for Cell Science, Pune and maintained in Dulbeccos Modified Eagle Medium (DMEM) containing high glucose with 10% fetal bovine serum (FBS) and supplemented with penicillin (5 units/mL) and streptomycin (5 μg/mL) in 5% CO 2 incubator at 37 °C.

Techniques: