murine 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

Journal: Bioactive Materials

Article Title: Anisotropic mechanotransductive tissue constructs via brush-assisted bioprinting of microfiber-reinforced composite bioinks

doi: 10.1016/j.bioactmat.2025.12.017

Figure Lengend Snippet: Comparative analysis of conventional and brush-assisted bioprinting on cellular behavior. (a) Schematic illustration of shear stress distribution in normal versus brush-assisted printing. (b) Overview of the brush-assisted printing setup. (c) SEM images of collagen fibrils and fluorescence images showing cell viability (live/dead) and cytoskeletal organization (DAPI/phalloidin) of C2C12 myoblasts. Quantification of (d) cell viability post-printing (n = 4), (e) cell metabolic activity (MTT assay, in situ /day 3/day 7, n = 4), (f) nuclei aspect ratio (n = 180), (g) orientation factor (n = 3), and (h) F-actin–positive area at day 3 (n = 10). (i) Comparing normal and brush-assisted printing the mechanotransduction pathways activated by shear stress and collagen alignment. (j) Heatmap of relative gene expression ( YAP, TAZ, AKT1, PIEZO1, PI3K, and CAPN2 ) after 7 days of culture (n = 4). (k) Agarose gel electrophoresis of PCR products from cells cultured on normal versus brush-printed scaffolds for 7 days. (l) Schematic illustration of blocking mechano-sensing ion channel with GsMTx-4. (m) Relative gene expression levels associated with mechanosensing channel and ca 2+ pathway (n = 5), (n) Hippo pathway (n = 5), (o) PI3K-AKT pathway (n = 5). Student's t-test was applied for two-group comparisons, and one-way ANOVA with Tukey's HSD post-hoc test was used for multiple comparisons. Data are presented as mean ± standard deviation (SD). Statistical significance was set at p < 0.05 (∗), p < 0.01 (∗∗), and p < 0.001 (∗∗∗). Abbreviation: MTT, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (cell metabolic activity assay); F-actin, Filamentous actin; PI3K , Phosphoinositide 3-kinase; CAPN2 , Calcium-activated neutral protease 2.

Article Snippet: H9C2 cardiomyoblasts (Korean Cell Line Bank, Seoul, Korea) and C2C12 murine skeletal muscle myoblasts (CRL-1772, ATCC, Manassas, USA) were cultured in high-glucose DMEM (Welgene, Korea) supplemented with 10 % fetal bovine serum (FBS) and 1 % penicillin–streptomycin (PS).

Techniques: Shear, Fluorescence, Activity Assay, MTT Assay, In Situ, Gene Expression, Agarose Gel Electrophoresis, Cell Culture, Blocking Assay, Standard Deviation, Metabolic Assay

Journal: Bioactive Materials

Article Title: Anisotropic mechanotransductive tissue constructs via brush-assisted bioprinting of microfiber-reinforced composite bioinks

doi: 10.1016/j.bioactmat.2025.12.017

Figure Lengend Snippet: Physical and biological evaluation of bioconstructs reinforced with straight (CSP) and coiled (CCP) PCL fibers. (a) Schematic illustration and optical images of flow tests with Col (collagen-only), CSP, and CCP bioinks, in which droplets were tilted at 90° for 10 min. (b) Quantification of droplet outflow (n = 10). Rheological measurements of bioinks: (c) storage modulus (G′) from frequency sweep (n = 3), (d) G′ and G″ from temperature sweep (n = 3), and (e) G′ and G″ under cyclic stress loading (10 and 200 Pa) showing viscoelastic recovery (n = 3). (f) SEM images of CCP constructs highlighting coiled PCL fiber and aligned collagen fibrils. (g) Optical image, live/dead staining, DAPI/phalloidin staining, and orientation factor analysis of C2C12 cells and PCL microfibers. Quantification of (h) cell viability (live/dead, n = 4), (i) F-actin–positive area (n = 20), and (j) metabolic activity (MTT assay, in situ /day 3/day 7, n = 4). Student's t-test was applied for two-group comparisons, and one-way ANOVA with Tukey's HSD post-hoc test was used for multiple comparisons. Data are presented as mean ± standard deviation (SD). Statistical significance was set at p < 0.05 (∗), p < 0.01 (∗∗), and p < 0.001 (∗∗∗).

Article Snippet: H9C2 cardiomyoblasts (Korean Cell Line Bank, Seoul, Korea) and C2C12 murine skeletal muscle myoblasts (CRL-1772, ATCC, Manassas, USA) were cultured in high-glucose DMEM (Welgene, Korea) supplemented with 10 % fetal bovine serum (FBS) and 1 % penicillin–streptomycin (PS).

Techniques: Construct, Staining, Activity Assay, MTT Assay, In Situ, Standard Deviation

Journal: Bioactive Materials

Article Title: Anisotropic mechanotransductive tissue constructs via brush-assisted bioprinting of microfiber-reinforced composite bioinks

doi: 10.1016/j.bioactmat.2025.12.017

Figure Lengend Snippet: In vitro myogenic differentiation of C2C12 cells cultured on Col, CSP and CCP scaffolds. (a) Live/dead staining at in situ , DAPI/phalloidin staining at day 3, and DAPI/ MHC staining at day 14. (b) Quantification of cell viability from live/dead assays (n = 4). (c) Nuclei orientation factor after 7 days of culture (n = 4). (d) Nuclei aspect ratio (n = 20). (e) F-actin positive area (n = 4). (f) Quantification of MHC fusion index (left, n = 5) and MHC maturation rate (right, n = 5). (g) Relative gene expression analysis and (h) agarose gel electrophoresis regarding mechanotransduction-related genes ( CAPN2, PIEZO1, RhoA, YAP, and TAZ ) (n = 4). (i) Western blot analysis of PIEZO1 . (j) Schematic illustrating differentiation progression and major genes involved at each stage. (k) Heatmap and (l) agarose gel electrophoresis of PCR products showing relative expression of myogenic markers ( MYF5, MYOD1, MYOG, MHC, MYH2, and MYH4 ) after 21 days of culture (n = 4). (m) Western blot analysis of MHC . Student's t-test was applied for two-group comparisons, and one-way ANOVA with Tukey's HSD post-hoc test was used for multiple comparisons. Data are presented as mean ± standard deviation (SD). Statistical significance was set at p < 0.05 (∗), p < 0.01 (∗∗), and p < 0.001 (∗∗∗). Abbreviation: MYF5 , Myogenic factor 5; MYOD1 , Myogenic differentiation 1; MYOG , Myogenin; MHC , Myosin heavy chain; MYH2 , Myosin heavy chain 2; MYH4 , Myosin heavy chain 4.

Article Snippet: H9C2 cardiomyoblasts (Korean Cell Line Bank, Seoul, Korea) and C2C12 murine skeletal muscle myoblasts (CRL-1772, ATCC, Manassas, USA) were cultured in high-glucose DMEM (Welgene, Korea) supplemented with 10 % fetal bovine serum (FBS) and 1 % penicillin–streptomycin (PS).

Techniques: In Vitro, Cell Characterization, Cell Culture, Staining, In Situ, Gene Expression, Agarose Gel Electrophoresis, Western Blot, Expressing, Standard Deviation

Journal: Molecular Medicine Reports

Article Title: Veronicastrum sibiricum (L.) Pennell extract alleviates inflammation-induced muscle atrophy through NLRP3 inflammasome regulation and mitochondrial function restoration

doi: 10.3892/mmr.2026.13857

Figure Lengend Snippet: Effects of VSE on inflammation-induced muscle atrophy in C2C12 myotubes. All experiments were carried out after C2C12 myoblasts had differentiated into myotubes for 4 days. C2C12 myotubes were pretreated with 0 (LPS only), 1.25, 2.5 or 5 µg/ml VSE for 3 h and then co-treated with 500 ng/ml LPS for (B, D and E) 24 h and (C) 48 h. Controls received no treatments. (A) C2C12 myotubes were exposed to 0–10 µg/ml VSE for 48 h to assess cytotoxicity. (B) mRNA levels of muscle-specific E3 ubiquitin ligases MuRF1 and atrogin-1 were measured using reverse transcription-quantitative PCR. (C) Giemsa-stained images of C2C12 myotubes showing changes in relative fiber width after treatment. CON, untreated control; LPS, 500 ng/ml; 1.25, 2.5 and 5, co-treatment with LPS (500 ng/ml) and VSE at the indicated concentrations (µg/ml). Analysis was performed using ImageJ (National Institutes of Health). Scale bar, 100 µm. Western blot analysis of (D) MuRF1, MaFbx and MyHC protein levels, and (E) phosphorylated Akt and Foxo3a, including membrane images and semi-quantitative analysis using ImageJ. All data are presented as the mean ± SD of triplicate results and statistical analysis was carried out using one-way ANOVA with Tukey's post hoc test. ## P<0.01 and #### P<0.0001 vs. control group; *P < 0.05, **P < 0.01, *** P < 0.001, ****P < 0.0001 vs. LPS-treated group. MuRF1, muscle-specific RING finger protein 1; MaFbx, muscle atrophy F-box protein; MyHC, myosin heavy chain; CON, control; LPS, lipopolysaccharide; VSE, Veronicastrum sibiricum seed extract; p-, phosphorylated; ns, not significant.

Article Snippet: Murine C2C12 myoblast cells were obtained from American Type Culture Collection.

Techniques: Ubiquitin Proteomics, Reverse Transcription, Real-time Polymerase Chain Reaction, Staining, Control, Western Blot, Membrane

Journal: Molecular Medicine Reports

Article Title: Veronicastrum sibiricum (L.) Pennell extract alleviates inflammation-induced muscle atrophy through NLRP3 inflammasome regulation and mitochondrial function restoration

doi: 10.3892/mmr.2026.13857

Figure Lengend Snippet: VSE downregulates the NLRP3 inflammasome. C2C12 myotubes were pretreated with VSE at 0 (LPS only), 1.25, 2.5 and 5 µg/ml for 3 h and then co-treated with 500 ng/ml LPS for 24 h. Controls received no treatments. (A) mRNA expression levels of NLRP3 , the NLRP3 inflammasome initiator, were analyzed using reverse transcription-quantitative PCR. (B) Protein expression levels of factors associated with the NLPR3 pathway, a major pyroptosis pathway, were analyzed using western blotting and ImageJ. All data are presented as the mean ± SD of triplicate results and statistical analysis was carried out using one-way ANOVA with Tukey's post hoc test. #### P<0.0001, ### P<0.001, ## P<0.01 vs. control; *P<0.05, **P<0.01, ***P<0.001, ****P<0.0001 vs. LPS treatment. NLRP3, NLR family pyrin domain containing 3; GSDMD, gasdermin-D; LPS, lipopolysaccharide; VSE, Veronicastrum sibiricum seed extract.

Article Snippet: Murine C2C12 myoblast cells were obtained from American Type Culture Collection.

Techniques: Expressing, Reverse Transcription, Real-time Polymerase Chain Reaction, Western Blot, Control

Journal: Molecular Medicine Reports

Article Title: Veronicastrum sibiricum (L.) Pennell extract alleviates inflammation-induced muscle atrophy through NLRP3 inflammasome regulation and mitochondrial function restoration

doi: 10.3892/mmr.2026.13857

Figure Lengend Snippet: VSE reduces the secretion of proinflammatory cytokines. C2C12 myotubes were pretreated with VSE at 0 (LPS only), 1.25, 2.5 and 5 µg/ml for 3 h and then co-treated with 500 ng/ml LPS for 24 h. Controls received no treatments. (A) mRNA expression levels of the proinflammatory cytokines TNF-α and IL-1β were analyzed using reverse transcription-quantitative PCR. (B) Protein expression levels of the proinflammatory cytokines TNF-α and IL-1β in the supernatant of C2C12 myotube cultures were analyzed using ELISA. All data are presented as the mean ± SD of triplicate results and statistical analysis was carried out using one-way ANOVA with Tukey's post hoc test. #### P<0.0001 vs. control; ***P<0.001, ****P<0.0001 vs. LPS treatment. A450, absorbance at 450 nm; LPS, lipopolysaccharide; VSE, Veronicastrum sibiricum seed extract.

Article Snippet: Murine C2C12 myoblast cells were obtained from American Type Culture Collection.

Techniques: Expressing, Reverse Transcription, Real-time Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay, Control

Journal: Molecular Medicine Reports

Article Title: Veronicastrum sibiricum (L.) Pennell extract alleviates inflammation-induced muscle atrophy through NLRP3 inflammasome regulation and mitochondrial function restoration

doi: 10.3892/mmr.2026.13857

Figure Lengend Snippet: VSE exerts antioxidant effects in C2C12 myotubes. C2C12 myotubes were pretreated with 0 (LPS only), 1.25, 2.5 and 5 µg/ml VSE for 3 h and then co-treated with 500 ng/ml LPS for 24 h. (A) Protein expression levels of antioxidant-related factors HO-1, Nrf2 and Keap1, and mitochondrial metabolism-related factor PGC-1α were analyzed using western blotting. (B) Images (left) were obtained under a fluorescence microscope to investigate the generation of mitochondrial ROS using ImageJ (right). Scale bar, 100 µm. All data are presented as the mean ± SD of triplicate results, and statistical analysis was carried out using one-way ANOVA with Tukey's post hoc test. # P<0.05, ## P<0.01, #### P<0.0001 vs. control; *P<0.05, **P<0.01, ***P<0.001, ****P<0.0001 vs. LPS treatment. LPS, lipopolysaccharide; VSE, Veronicastrum sibiricum seed extract; ROS, reactive oxygen species; HO-1, heme oxygenase-1; Nrf2, nuclear factor erythroid 2-related factor 2; Keap1, kelch like ECH associated protein 1; PGC-1α, peroxisome proliferator-activated receptor γ coactivator 1α; CON, control.

Article Snippet: Murine C2C12 myoblast cells were obtained from American Type Culture Collection.

Techniques: Expressing, Western Blot, Fluorescence, Microscopy, Control

Journal: Molecular Medicine Reports

Article Title: Veronicastrum sibiricum (L.) Pennell extract alleviates inflammation-induced muscle atrophy through NLRP3 inflammasome regulation and mitochondrial function restoration

doi: 10.3892/mmr.2026.13857

Figure Lengend Snippet: VSE exerts protective effects on mitochondrial function, reducing functional impairment under inflammatory conditions. C2C12 myotubes were pretreated with 0 (LPS only), 1.25, 2.5 and 5 µg/ml VSE for 3 h and then co-treated with 500 ng/ml LPS for 24 h. (A) Protein expression levels of the mitochondrial dynamics-related OXPHOS genes, including CI-CV, were analyzed using western blotting. (B) JC-1 staining was carried out to assess the mitochondrial membrane potential. Representative images of JC-1 fluorescence (red/green) were analyzed using ImageJ (National Institutes of Health). Scale bar, 25 µm. All data are presented as the mean ± SD of triplicate results and statistical analysis was carried out using one-way ANOVA with Tukey's post hoc test. ## P<0.01, #### P<0.0001 vs. control; *P<0.05, **P<0.01, ***P<0.001, ****P<0.0001 vs. LPS treatment. LPS, lipopolysaccharide; VSE, Veronicastrum sibiricum seed extract; CI-CV, complexes I–V; OXPHOS, oxidative phosphorylation system; CON, control.

Article Snippet: Murine C2C12 myoblast cells were obtained from American Type Culture Collection.

Techniques: Functional Assay, Expressing, Western Blot, Staining, Membrane, Fluorescence, Control, Phospho-proteomics

Journal: Physiological Reports

Article Title: Synthetic estrogen and progestin effects on the myogenic program following damage in C2C12 murine myoblasts

doi: 10.14814/phy2.70886

Figure Lengend Snippet: (a) Representative images of C2C12 myotubes 6 days after differentiation in vehicle (VEH), OC1, OC2, OC3, and OC4 formulations. Myotubes were stained with desmin (green) and DAPI (blue). (b, d, f) Myotube diameter (um) of vehicle, EE, progestin, and OC formulations after 6 days of differentiation. (c, e, g) Myonuclear index of vehicle, EE, progestin and OC formulations after 6 days of differentiation. Values are presented as median lines and interquartile range (boxes) ± maximum and minimum values (whiskers), with + representing the mean. Statistical comparisons were performed using a one‐way ANOVA. p ‐value indicates a significant difference from vehicle (VEH).

Article Snippet: Murine C2C12 skeletal muscle myoblasts from American Type Culture Collection (Cedarlane) were cultured in uncoated 150 mm tissue culture dishes in 5% CO 2 at 37°C.

Techniques: Staining

Journal: bioRxiv

Article Title: C26 and CT26 colorectal cancer models exhibit divergent cachexia phenotypes, intramuscular inflammation, and protein turnover signaling

doi: 10.64898/2026.04.21.719997

Figure Lengend Snippet: (A) Subconfluent C2C12 myoblasts cultured in DMEM were plated in the 24-well base plate at 2.5 × 10 4 /cm 2 . C2C12 myoblasts proliferated for four days and were induced to differentiate in differentiation media (DM), DMEM containing 2% horse serum for 72 hours. On the same day of inducing differentiation, C26 cells and CT26 cells were plated in the co-culture inserts at increasing densities (4.17 × 10 4 /cm 2 , 8.3 × 10 4 /cm 2 , and 1.67 × 10 5 /cm 2 ) and proliferated for 72 hours. C2C12 myoblasts were plated in the inserts at 8.3 × 10 4 /cm 2 as a negative co-culture control. Then, the co-culture inserts containing C26, CT26, or C2C12 cells were moved to the lower compartments of the base plate. Fresh DM was added to both inserts and lower compartments. (B) The co-culture continued for 72 hours, and cells were fixed with 4% paraformaldehyde (PFA) and stained for myosin heavy chain (MyHC, green) and myogenin (MyoG, red). DAPI was used to counterstain the nuclei (blue). Scale bar is set at 200 µm. Myotube diameter (C) , percentage of myotube area (D) , MyoG + /total DAPI + percentage (E) , and total DAPI + cells (F) were quantified to assess potential atrophic effect of C26 cells and CT26 cells. *P<0.05 for pairwise comparison to C2C12 insert control, #P<0.05 for comparison between C26 and CT26 cells.

Article Snippet: The murine skeletal muscle cell line C2C12 (ATCC, CRL-1772) and two colorectal carcinoma cell lines including C26 (a gift from Dr. Keehong Kim at Purdue University) and CT26 (ATCC, CRL-2638) were cultured separately in Dulbecco’s modified Eagle medium (DMEM, Gibco, 11995-065) containing 10% fetal bovine serum (FBS, Corning, 35-010-CV) and antibiotics (penicillin 100 U/mL, streptomycin 100 μg/mL, Gibco, 15140-122) at 37 °C in a cell incubator with 5% CO 2 .

Techniques: Cell Culture, Co-Culture Assay, Control, Staining, Comparison