mouse muscle cell line c2c12  (ATCC)

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