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: 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

Journal: RSC Chemical Biology

Article Title: Signatures of native-like glycosylation in RNA replicon-derived HIV-1 immunogens

doi: 10.1039/d5cb00165j

Figure Lengend Snippet: Differential glycan composition on Env expressed in different cell lines. Glycan composition at reporter glycosylation sites predominantly displaying: oligomannose-type glycans (N332), a mixture of complex- and oligomannose-type glycans (N355), and complex-type glycans (N88). Four different production systems were used as described in the key, including expression in three different cell lines, HEK293F, C2C12 and DC2.4 cells. All the glycan composition observed in the site-specific analysis are shown on the left, simplified in distinct categories represented as, core, oligomannose-type, and complex-type glycans. Some compositions annotated as complex-type glycans can exhibit isomers formally constituting hybrid-type glycans. Represented glycan compositions (See Methods for full classification) are colored according to the scale provided in the right for each category of glycan composition. The representative glycan composition data at all the sites is the average of three or more biological replicates.

Article Snippet: The mouse muscle cell line (C2C12) was cultured in Dulbecco's modified Eagles medium supplemented with 10% fetal bovine serum (FBS) as recommended in manufacturer's protocol (American type culture collection, Catalogue no. CRL-1772).

Techniques: Glycoproteomics, Expressing

Journal: iScience

Article Title: Proteomics- and BRET- screens identify SPRY2 as a Ras effector that impacts its membrane organization

doi: 10.1016/j.isci.2025.113974

Figure Lengend Snippet: SPRY2 is recruited to active K-Ras on the plasma membrane (A) AlphaFold 3 prediction of K-Ras/SPRY2 complex, with zoom-in showing residues with putative side chain interactions. (B and C) BRET-titration curves of the nL-tagged K-RasG12V or K-RasG12V-Q25A/Y40A/R41A (mutK-RasG12V) interactions with mNG-tagged SPRY2 or SPRY2-K187A/Y191A/T298A (mutSPRY2) (B), respectively corresponding C-SPRY2 variants (C) in HEK cells from N = 2-4 independent biological repeats. Means ± SEM of BRETtop were analyzed using One-Way Brown-Forsythe and Welch ANOVA tests with Dunnett's T3 correction for multiple comparisons. (D) AlphaFold 3 prediction of the SPRY2/SPRY2 interaction, with zoom-in showing residues within 3 Å distance. (E) BRET-titration curves of the nL-SPRY2/mNG-SPRY2 and nL-SPRY2/mNG-SPRY4 interaction in HEK cells with means ± SEM of BRETtop from N = 4 independent biological repeats. (F) BRET-titration curve of the wt or mutant nL-K-RasG12V with mNG-SPRY4 in HEK cells with means ± SEM of BRETtop from N = 3–6 independent biological repeats. (G) Flow cytometric quantification of MyHC terminal differentiation marker expression in C2C12 cells in low serum for 72 h after transfection with mNG-SPRY2, mNG-N-SPRY2, or mNG-C-SPRY2 constructs. Means ± SD are plotted from N = 3 biological repeats. Statistical analysis was done using one-way ANOVA and Dunn’s post hoc test. (H) Schematic illustrating our speculative models for how APLP2 (left) and SPRY2 (right) impact Ras membrane organization and activity. See the main text for more details. See also and .

Article Snippet: The mouse muscle C2C12 cell line (female mouse origin) was purchased from ATCC (CRL-1772) and cultured in DMEM supplemented with ∼9% (v/v) FBS, 2 mM L- and penicillin-streptomycin 10,000 units/mL (high serum medium).

Techniques: Clinical Proteomics, Membrane, Titration, Mutagenesis, Marker, Expressing, Transfection, Construct, Activity Assay