Journal: Biological Research

Article Title: MicroRNA-721 regulates gluconeogenesis via KDM2A-mediated epigenetic modulation in diet-induced insulin resistance in C57BL/6J mice

doi: 10.1186/s40659-024-00495-0

Figure Lengend Snippet: MiR-721 is upregulated under insulin resistant conditions. The figure shows volcano plot derived from GEO2R analysis of GEO dataset GSE94799 ( A ), miR-721 expression in diet induced insulin resistant mice ( B ) and miR-721 expression in palmitate induced insulin resistant Hepa 1–6 cells ( C ). Data are expressed as mean ± SD, (n = 3) ##p < 0.01, ###p < 0.001, (#) vs CON

Article Snippet: Hepa 1–6 (Murine hepatoma cell line) was acquired from National Centre for Cell Sciences (NCCS) Pune, India.

Techniques: Derivative Assay, Expressing

Journal: Biological Research

Article Title: MicroRNA-721 regulates gluconeogenesis via KDM2A-mediated epigenetic modulation in diet-induced insulin resistance in C57BL/6J mice

doi: 10.1186/s40659-024-00495-0

Figure Lengend Snippet: miR-721 enhances glucose production in Hepa 1–6 cells. Cells were transfected with miR-721 mimic or treated with palmitate for 24 h and glucose production was evaluated. The figure shows glucose production ( A , E ), expression of FOXO1 ( B , F ), PCK1 ( C , G ) and G6PC ( D , H ) in response to miR-721 mimic and palmitate. Data are expressed as mean ± SD, (n = 3) #p < 0.05, ##p < 0.01, ###p < 0.001, (#) vs CON. (SM-Scramble mimic, MM-miR-721 mimic)

Article Snippet: Hepa 1–6 (Murine hepatoma cell line) was acquired from National Centre for Cell Sciences (NCCS) Pune, India.

Techniques: Transfection, Expressing

Journal: Biological Research

Article Title: MicroRNA-721 regulates gluconeogenesis via KDM2A-mediated epigenetic modulation in diet-induced insulin resistance in C57BL/6J mice

doi: 10.1186/s40659-024-00495-0

Figure Lengend Snippet: miR-721 affects gluconeogenesis via KDM2A/FOXO1 pathway. Bioinformatic analysis identified KDM2A as a potential target and to further validate the effects of miR-721 inhibition on KDM2A, Hepa 1–6 cells were transfected with miR-721 inhibitor and treated with daminozide a KDM2A inhibitor and effects on glucose production were evaluated. The figure shows the binding pose and binding free energy for miR-721 and KDM2A mRNA predicted by StarMir ( A ), mRNA expression of Kdm2a ( B ), glucose production ( C ), mRNA expression of Foxo1, Pck1, G6pc ( D – F ), immunoblots and quantification of KDM2A, FOXO1, PCK1, G6PC (G-K) and CHIP-qPCR of H3K36me2 for enrichment on FOXO1 promoter (L). Data are expressed as mean ± SD, (n = 3) *p < 0.05, ## or **p < 0.01, ### or ***p < 0.001, (#) vs CON and (*) vs PAL + SI group (SI: Scramble inhibitor and MI: miR-721 inhibitor)

Article Snippet: Hepa 1–6 (Murine hepatoma cell line) was acquired from National Centre for Cell Sciences (NCCS) Pune, India.

Techniques: Inhibition, Transfection, Binding Assay, Expressing, Western Blot

Journal: Biochemical and biophysical research communications

Article Title: Ablation of Dicer leads to widespread perturbation of signaling pathways

doi: 10.1016/j.bbrc.2015.05.077

Figure Lengend Snippet: Investigation of phosphotyrosine profile upon ablation of Dicer A) Phosphotyrosine profile of control and Dicer knockout mice. Ctrl1 and Ctrl2 represent liver lysates from two control mice and KO 1 and KO 2 represent liver lysates from 2 Dicer knockout mice. The lysates were immunoblotted using anti-phosphotyrosine antibody. B) Schematic representation of workflow followed to identify differentially phosphorylated tyrosine sites upon ablation of Dicer. Pooled liver lysates from 5 control and 5 Dicer knockout mice were spiked with 13C6-lysine and 13C6-ariginine labeled Hepa 1–6 cell lysates and replicate analysis was carried out. Phosphotyrosine containing peptides were enriched using pY100 anti-phosphotyrosine antibody. LC-MS/MS analysis was carried out using LTQ-Orbitrap Velos followed by data analysis.

Article Snippet: Mouse hepatoma Hepa 1–6 cells were (kind gift from Kensler Lab, School of Public Health, Johns Hopkins University) adapted to 13 C 6 -Lysine; 13 C 6 -Arginine containing custom DMEM media to label proteins in vivo .

Techniques: Control, Knock-Out, Labeling, Liquid Chromatography with Mass Spectroscopy