c2c12 myoblasts  (ATCC)

Journal: Journal of Enzyme Inhibition and Medicinal Chemistry

Article Title: Flavonoids from Elsholtzia ciliata restore redox electron flow and metabolic signaling via PTP1B inhibition in muscle and liver cells

doi: 10.1080/14756366.2026.2666369

Figure Lengend Snippet: Principal Component Analysis (PCA) biplots visualise the distinct mechanistic clustering of E. ciliata -derived compounds and reference drugs across all in vitro models. PCA biplots were generated from the combined functional (glucose uptake) and gene expression (qRT-PCR) data for each of the four models: (A) C2C12-Insulin, (B) C2C12-Leptin, (C) C2C12-Adiponectin, and (D) Hepa1c1c7-PA con. Data points (x) represent individual biological replicates, and the ellipses represent the 95% confidence intervals for each treatment group. Loading vectors (black arrows) indicate the direction and magnitude of influence for each variable (e.g. Ptpn1 , Ampka1 , Glucose uptake) on the principal components (PC1 and PC2). The percentage of variance explained by each principal component is shown on the axes. The plots visually confirm the clear separation of the disease control groups (PA-treated) and the distinct mechanistic clusters formed by EC2, EC5, Met, and NAC.

Article Snippet: Murine C2C12 myoblasts (ATCC, Manassas, VA, USA) were cultured in Dulbecco’s Modified Eagle’s Medium (DMEM) supplemented with 10% foetal bovine serum (FBS; Gibco, Thermo Fisher Scientific, Waltham, MA, USA) and 1% penicillin–streptomycin (Gibco) .

Techniques: Derivative Assay, In Vitro, Generated, Functional Assay, Gene Expression, Quantitative RT-PCR, Control

Journal: Journal of Enzyme Inhibition and Medicinal Chemistry

Article Title: Flavonoids from Elsholtzia ciliata restore redox electron flow and metabolic signaling via PTP1B inhibition in muscle and liver cells

doi: 10.1080/14756366.2026.2666369

Figure Lengend Snippet: Baseline glucose uptake and metabolic gene expression in differentiated C2C12 myotubes under non-PA conditions. Differentiated C2C12 myotubes were treated for 24 h with each of metabolic hormones (insulin, leptin, or adiponectin), reference antidiabetic agents (Met, Duo, NAC), or E. ciliata -derived compounds (EC2–EC5). (A) Glucose uptake (%) was measured in hormone/antidiabetic agent-treated cells (top) and EC-treated cells (bottom). (B) Relative levels of gene expression of Ptpn1 , Pparg , Ampka1 , Ampka2 , and Txnip was assessed via qRT-PCR. ECs were treated at the higher dose (20 μM). The concentration of ECs used in this experiment was 10 μM (L) or 20 μM (H). * p < 0.05, ** p < 0.01, *** p < 0.001 vs. non-treated (vehicle)control. PA , palmitate; Ins-C , insulin- control; Lep-C, leptin-control; Adi-C, Adiponectin-control ; ScI, sc-222227; UA, ursolic acid; Met, metformin; Duo, Duoglow (pioglitazone + dapagliflozin); NAC, N-acetylcysteine; EC2–EC5, E. ciliata -derived compounds; Ptpn1 , protein tyrosine phosphatase 1B; Pparg , peroxisome proliferator-activated receptor gamma; Ampka1/2 , AMP-activated protein kinase alpha 1/2; Glut4 , glucose transporter 4.

Article Snippet: Murine C2C12 myoblasts (ATCC, Manassas, VA, USA) were cultured in Dulbecco’s Modified Eagle’s Medium (DMEM) supplemented with 10% foetal bovine serum (FBS; Gibco, Thermo Fisher Scientific, Waltham, MA, USA) and 1% penicillin–streptomycin (Gibco) .

Techniques: Gene Expression, Derivative Assay, Quantitative RT-PCR, Concentration Assay, Control

Journal: Journal of Enzyme Inhibition and Medicinal Chemistry

Article Title: Flavonoids from Elsholtzia ciliata restore redox electron flow and metabolic signaling via PTP1B inhibition in muscle and liver cells

doi: 10.1080/14756366.2026.2666369

Figure Lengend Snippet: Restoration of the impairment of glucose uptake by E. ciliata -derived compounds in PA-induced insulin-resistant C2C12 myotubes. (A) Glucose uptake levels (%) were quantified. (B) Relative levels of mRNA expression of Ptpn1 , Glut4 , Pi3k , and Akt were analysed via qRT-PCR. EC2-H and EC5-H downregulated Ptpn1 and restored Glut4 levels. ECs were treated at the dose of 20 μM. The concentration of ECs used in this experiment was 10 μM (L) or 20 μM (H). Data are presented as mean ± SEM ( n = 3). * p < 0.05, ** p < 0.01 vs. Insulin + PA control group. † p < 0.05, †† p < 0.01 vs. insulin control group. PA, palmitate; Ins-C, insulin-only control; Ins-PA, insulin + PA; ScI, sc-222227; UA, ursolic acid; Met, metformin; Duo, Duoglow (pioglitazone + dapagliflozin); NAC, N-acetylcysteine; EC2–EC5, E. ciliata -derived compounds; Ptpn1 , protein tyrosine phosphatase 1B; Glut4 , glucose transporter 4; Pi3k , phosphoinositide 3-kinase; Akt , protein kinase B.

Article Snippet: Murine C2C12 myoblasts (ATCC, Manassas, VA, USA) were cultured in Dulbecco’s Modified Eagle’s Medium (DMEM) supplemented with 10% foetal bovine serum (FBS; Gibco, Thermo Fisher Scientific, Waltham, MA, USA) and 1% penicillin–streptomycin (Gibco) .

Techniques: Derivative Assay, Expressing, Quantitative RT-PCR, Concentration Assay, Control

Journal: Journal of Enzyme Inhibition and Medicinal Chemistry

Article Title: Flavonoids from Elsholtzia ciliata restore redox electron flow and metabolic signaling via PTP1B inhibition in muscle and liver cells

doi: 10.1080/14756366.2026.2666369

Figure Lengend Snippet: Reversal of leptin resistance by E. ciliata -derived compounds in PA-treated C2C12 myotubes. Glucose uptake in response to leptin stimulation. Glucose uptake was expressed as a percentage of normal cells. (B) Gene expression analysis of Ptpn1 , Jak2 , Stat3 , and Glut4 was performed by qRT-PCR. ECs were treated at the dose of 20 μM. Data are presented as mean ± SEM ( n = 3). * p < 0.05, ** p < 0.01 vs. Lep + PA control group. † p < 0.05, †† p < 0.01 vs. Leptin control group. PA, palmitate; Lep-C, leptin-only control; Lep-PA, leptin + PA; ScI, sc-222227; UA, ursolic acid; Met, metformin; Duo, Duoglow (pioglitazone + dapagliflozin); NAC, N-acetylcysteine; EC2–EC5, E. ciliata -derived compounds; Ptpn1 , protein tyrosine phosphatase 1B; Jak2 , Janus kinase 2; Stat3 , signal transducer and activator of transcription 3; Glut4 , glucose transporter 4.

Article Snippet: Murine C2C12 myoblasts (ATCC, Manassas, VA, USA) were cultured in Dulbecco’s Modified Eagle’s Medium (DMEM) supplemented with 10% foetal bovine serum (FBS; Gibco, Thermo Fisher Scientific, Waltham, MA, USA) and 1% penicillin–streptomycin (Gibco) .

Techniques: Derivative Assay, Gene Expression, Quantitative RT-PCR, Control

Journal: Journal of Enzyme Inhibition and Medicinal Chemistry

Article Title: Flavonoids from Elsholtzia ciliata restore redox electron flow and metabolic signaling via PTP1B inhibition in muscle and liver cells

doi: 10.1080/14756366.2026.2666369

Figure Lengend Snippet: Effects of E. ciliata -derived compounds on redox homeostasis: NAD⁺/NADH, NADP⁺/NADPH ratios, and FAD levels. C2C12 myotubes were co-treated with palmitate (PA, 300 μM for 48 h) and each test compound or hormone for the final 24 h. The NAD⁺/NADH ratio (A), NADP⁺/NADPH ratio (B), and FAD (C) levels were measured to assess redox homeostasis. Data are presented as mean ± SEM ( n = 5). * p < 0.05, ** p < 0.01 vs. corresponding –PA control group (i.e. basal); † p < 0.05, †† p < 0.01 vs. PA-treated (vehicle) control group. PA, palmitate; ScI, sc-222227; UA, ursolic acid; Met, metformin; Duo, Duoglow (pioglitazone + dapagliflozin); NAC, N-acetylcysteine; EC2–EC5, E. ciliata -derived compounds; NAD⁺, nicotinamide adenine dinucleotide (oxidised form); NADH, nicotinamide adenine dinucleotide (reduced form); NADP⁺, nicotinamide adenine dinucleotide phosphate (oxidised form); NADPH, nicotinamide adenine dinucleotide phosphate (reduced form); FAD, flavin adenine dinucleotide.

Article Snippet: Murine C2C12 myoblasts (ATCC, Manassas, VA, USA) were cultured in Dulbecco’s Modified Eagle’s Medium (DMEM) supplemented with 10% foetal bovine serum (FBS; Gibco, Thermo Fisher Scientific, Waltham, MA, USA) and 1% penicillin–streptomycin (Gibco) .

Techniques: Derivative Assay, Control

Journal: bioRxiv

Article Title: Pancreatic cancer extracellular vesicles carry a time-of-day-regulated miRNA cargo that disrupts the skeletal muscle clock and bioenergetics

doi: 10.64898/2026.05.03.722338

Figure Lengend Snippet: (A) Bioluminescence recording, ( B ) period analysis, and ( C ) phase and amplitude analysis of U2OS BMAL1 :Luc reporter cells treated with PANC-1 CM at 12.5%, 25%, 50%, and 100% of the recording media. ( D ) Bioluminescence recording, ( E ) period analysis, and ( F ) phase and amplitude analysis of NIH3T3 Bmal1 :Luc reporter cells treated with PANC-1 CM at the same concentrations. For all bioluminescence experiments, at least three complete oscillations were included in the period estimation, excluding the first 24 h. Mean ± SD of relative mRNA expression of NIH3T3 core clock genes Bmal1 ( G ), Per2 ( H ), and Cry2 ( I ) measured over 36 h in response to PANC-1 CM. Relative mRNA levels of core clock genes in synchronized C2C12 myotubes over 32 h following treatment with PANC-1 CM: ( J ) Bmal1 , ( K ) Per2 , and ( L ) Cry2 . Cosine curves were fit for visualization purposes only; solid lines represent rhythmic oscillations (p<0.05) detected by MetaCycle, while dashed lines indicate loss of statistical rhythmicity (Suppl. Table 1). ( G–L ) Black: Control; ( G–I ) Red: PANC-1 CM; ( J–L ) Blue: PANC-1 CM. ( M ) Schematic representation and representative images of mature C2C12 myotube atrophy in response to NIH3T3 or PANC-1 released factors using a Transwell co-culture system; three measurements per myotube (yellow arrows) were used to quantify shortening. ( N ) Quantification of normalized myotube diameter under NIH3T3 vs PANC-1 co-culture, normalized to NIH3T3 co-culture control. One-way ANOVA: ( B ) p=0.0009, ( E ) p=0.0087. ( B, E ) Dunnett’s post-hoc test: *p<0.05; **p<0.01; ***p<0.001. (N) Student’s t-test: ***p<0.001.

Article Snippet: The human pancreatic cancer cell line PANC-1, the murine fibroblast cell line NIH3T3, and the murine myoblast cell line C2C12 were purchased from American Type Culture Collection (ATCC, Manassas, VA).

Techniques: Expressing, Control, Co-Culture Assay

Journal: bioRxiv

Article Title: Pancreatic cancer extracellular vesicles carry a time-of-day-regulated miRNA cargo that disrupts the skeletal muscle clock and bioenergetics

doi: 10.64898/2026.05.03.722338

Figure Lengend Snippet: (A) Top 35 miRNAs by mean expression in PANC-1-derived sEVs. Bar plot of mean log2 CPM across 9 time-points (4–36 h). Red bars: miRNAs selected from the top 35 to be tested in the BMAL1 :Luc reporter and atrophy assays; grey bars: remaining top-35 miRNAs. miRNAs are ranked in descending order of EV expression. ( B ) GO Biological Process enrichment of the experimentally validated targets (miRTarBase) of the 11 selected miRNAs. Terms are grouped into functional categories. Dot size represents the number of validated target genes associated with each term; dot color indicates Gene Ratio (proportion of input genes annotated to the term), from light pink (low) to dark red (high). Analysis performed with clusterProfiler. ( C , top panel) Normalized C2C12 myotube diameter at 0, 24, and 48 h post-transfection with miR-27b-3p, miR-615-3p, miR-191-5p, miR-127-3p, miR-99b-5p, or negative-transfection control (NTC); dexamethasone (Dexa) included as positive control. ( C , lower panel) Normalized C2C12 myotube diameter at the same time-points after transfection with hsa-let-7f-5p, miR-183-5p, miR-92a-3p, miR-30c-5p, miR-26a-5p, miR-10a-5p, NTC, or Dexa. ( D ) Oxygen consumption rate (OCR; top), resting-phenotype plot of basal OCR vs ECAR (middle), and metabolic-capacity plot of maximal OCR vs ECAR following FCCP (lower) for mature C2C12 myotubes 48 h post-transfection with miR-27b-3p, miR-615-3p, miR-191-5p, or NTC (Control). ( E ) Same panels for myotubes transfected with miR-127-3p, miR-99b-5p, miR-183-5p, or NTC. Sequential injections of oligomycin, FCCP, and rotenone/antimycin A were used to dissect mitochondrial respiration. Data are presented as mean ± SEM. ( C ) Measurements were taken from at least 5 random fields per well in N=3 wells; statistical analysis used 2-way ANOVA with Dunnett’s post-hoc correction: *p<0.05; **p<0.01; ***p<0.001; ****p<0.0001.

Article Snippet: The human pancreatic cancer cell line PANC-1, the murine fibroblast cell line NIH3T3, and the murine myoblast cell line C2C12 were purchased from American Type Culture Collection (ATCC, Manassas, VA).

Techniques: Expressing, Derivative Assay, Functional Assay, Transfection, Control, Positive Control