Journal: bioRxiv

Article Title: Increased utrophin expression in healthy and DMD patient derived myoblasts in response to ERK1/2 and EZH2 inhibitor treatment

doi: 10.64898/2026.04.13.718206

Figure Lengend Snippet: Corresponding to . a . Basal levels of MYOD1 mRNA in primary healthy human myoblasts (n=6) and primary DMD patient-derived myoblasts (n=16). Lines show median and quartiles. b . AB1098 cells were treated with 10 μM LY32 +/- 10 μM GSK503 or vehicle (DMSO) for 24 hours. Directly following treatment (0 days since drug removal) cells were manually counted, with trypan blue staining to discount dead cells. Cells were then replated at a constant concentration, cultured in proliferation media without drugs for 24 hours and counted again (1 day since drug removal). Cells were then replated at a constant concentration and cultured again for 6 days before counting (7 days since drug removal). Cells were passaged once in this 6-day period. Cell count is shown relative to the vehicle only control for each time point. c . RT-qPCR of MYOD1 in immortalised myoblasts AB1190 and AB1098 following treatment with varying concentrations of Ruxolitinib (Ruxo). Results are shown relative to vehicle treated sample with a two-way ANOVA with Dunnetts multiple comparison test to determine significance. Graph shows mean (n=3) +/- SEM. a-b . MYOD1 is normalised against TBP . *p<0.05, ***p<0.001, ****p<0.0001.

Article Snippet: The muscle biopsy was enzymatically digested in basal medium (C-23060; PromoCell, Heidelberg, Germany) containing collagenase D (2 mg/mL, Roche, Germany) and Dispase II (2 mg/mL, Sigma) for 1 h at 37°C with trituration every 15 min. After filtering through a 100-μm filter (BD Falcon) and centrifugation cells were resuspended in proliferation medium (15% FBS, C-23060; PromoCell, Heidelberg, Germany) and transferred to a T-25 tissue culture vessel (Nunc, Germany).

Techniques: Derivative Assay, Staining, Concentration Assay, Cell Culture, Cell Characterization, Control, Quantitative RT-PCR, Comparison

Journal: Scientific Reports

Article Title: The C16orf87 protein is a subunit of the MIER corepressor complex controlling embryonic development and cell migration

doi: 10.1038/s41598-026-50740-7

Figure Lengend Snippet: Knockdown of C16orf87 causes minor changes in the host cell protein profile. ( A ) Alignment of human ( Homo sapiens , UniProtKB accession number Q6PH81 ), mouse ( Mus musculus , UniProtKB accession number Q9CR55 ), and zebrafish ( Danio rerio , UniProtKB accession number Q6DGQ4 ) C16orf87 amino acid sequences. Alignment mismatches are indicated in gray boxes. The underlined sequence represents a possible minimal Akt/PKB kinase consensus recognition motif. A Ser91(S91) phosphorylation site is marked with an asterisk. ( B ) Per-residue confidence (pLDDT) coloring of the top-ranked predicted model of C16orf87. In the inset, the predicted zinc-ribbon domain is shown with the zinc-interacting cysteines (Cys16, Cys19, Cys30, and Cys32) indicated around the zinc ion (Zn 2+ ). The position of the phosphorylated serine (Ser91), a putative alpha-helix between amino acid residues Ser-107 and Ala-126, and the confidently predicted C-terminal alpha-helix between amino acid residues Asp-130 and Ile-153 are also highlighted. The ipTM and pTM values are annotated. N, N-terminus; C, C-terminus. Figure was rendered using ChimeraX (version 1.8, https://www.rbvi.ucsf.edu/chimerax ) ( C ) Western blot (WB) analysis of C16orf87 siRNA (siC16) knockdown in Panc-01, MiaPaCa-2, and C2C12 cell lines. A non-specific, scrambled siRNA (siScr) was used as a control; the WB membrane was probed with the antibodies against C16orf87 and actin. MS-based proteomics analysis of siRNA-treated C2C12 ( D ), MiaPaCa-2 ( E ), and Panc-01 ( F ) cells. Data points corresponding to histones are colored in pink, and statistically significant ( P < 0.05, fold-change > 1) proteins are colored in yellow (mouse cell line C2C12) and green (human cell lines, Panc-01 and MiaPaCa-2).

Article Snippet: Human pancreatic cancer cell lines Panc-01 (ATCC, CRL-1469) and MiaPaCa-2 (ATCC, CRL-1420), mouse skeletal muscle cell line C2C12 (ATCC, CRL-1772), and human cervical cancer cell line HeLa S3 (ATCC, CCL-2.2) were used in this study.

Techniques: Knockdown, Sequencing, Phospho-proteomics, Residue, Western Blot, Control, Membrane

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

Journal: bioRxiv

Article Title: Aspirin hastens resolution of skeletal muscle inflammation and promotes recovery of muscle strength following acute injury

doi: 10.64898/2026.04.21.719989

Figure Lengend Snippet: A: Representative immunofluorescence images of day 3 post-differentiation C2C12 myotubes that were treated during differentiation with various NSAIDs, including aspirin (ASA), indomethacin (INDO), ibuprofen (IBU), celecoxib, and SC-236 at the indicated concentrations. Cells were stained for myosin heavy chain (MyHC, green), myogenin (MyoG, red), and DAPI (nuclei, blue). B: Representative immunofluorescence images of day 3 post-differentiation C2C12 myotubes treated with doses of INDO ranging from 6.25 µM to 400 µM. C-E : Quantitative analysis of INDO-treated cells showing a dose-dependent decrease in DAPI + cell density ( C ), differentiation index (%) ( D ), and myotube area (%) ( E ). F: Representative images of ASA-treated myotubes ranging from 62.5 µM to 4 mM. H–J: Quantification of DAPI + cell density ( H ), differentiation index (%) ( I ), and myotube area (%) ( J ) in C2C12 cells receiving ASA treatment. K: Representative images comparing the effects of ASA (2 mM), INDO (200 µM), and the combination of INDO + ASA on C2C12 myotube morphology. L-N: Statistical comparison of cell density ( L ), differentiation index (%) ( M ), and myotube area (%) ( N ) across vehicle and treatment groups. Data are expressed as mean ± SEM. Statistical significance was determined by one-way ANOVA followed by Holm-Šídák post hoc tests. *, **, ***, and **** denotes p<0.05, p<0.01, p<0.001, and p<0.0001 between indicated groups, respectively. # denotes p<0.05 difference between ASA and vehicle groups.

Article Snippet: The mouse C2C12 skeletal muscle cell line (ATCC, CRL-1772) was cultured in high glucose Dulbecco’s modified Eagle medium (DMEM, Gibco, 11995-065) supplemented with 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 5% CO atmosphere.

Techniques: Immunofluorescence, Staining, Comparison