Goat anti mouse alexa fluor 488 ab150113

For immunofluorescence studies, cells grown on a 4-well plate were fixed with freshly prepared 4% paraformaldehyde in PBS for 15 min at room temperature and permeabilized and blocked with 1% Triton-X, 1% BSA, and 10% FBS-contained PBS for 1 h at room temperature. The cells were incubated with primary antibodies in 10% FBS, 1% BSA, and 1% Triton-X-containing PBS overnight at 4°C. On the following day, the cells were washed three times in 0.1% Triton/PBS and incubated with fluorescent-labeled IgG (H + L) secondary antibodies (Abcam, Goat anti-mouse (Alexa Fluor 488): ab150113, (Alexa Fluor 594): ab150116, Goat anti-rabbit (Alexa Fluor 594: ab150080) at 1:500 dilutions in 1% FBS, 1% BSA, and 1% Triton-X-containing PBS for 2 h at room temperature. The cells were washed three times and filled with 0.1% Triton-containing PBS, and nuclei were counter-stained with DAPI (Sigma-Aldrich, D9542) for immunofluorescence. Primary antibodies used in this study include Anti-Myosin Heavy Chain antibody (MF20) (1:500, R&D Systems, MAB4470), Anti-Myogenin antibody (1:500, Abcam, ab124800), Anti-p-Histone H3 antibody (C-2) (1:1000, Santa Cruz, sc-374669), and Anti-Cleaved Caspase-3 antibody (Asp175) (5A1E) (1:500, Cell Signaling Technology, #9664). The observation was performed under a microscope (Keyence, BZ-X710).

PCL (average MN 80,000) was obtained from Sigma Aldrich (USA). Dichloromethane (DCM) was purchased from Merck-Millipore (USA). C2C12 mouse myoblast cell line (CRL-1772™) was obtained from ATCC^® (UK). Dulbecco’s Modified Eagle Medium (DMEM, high and low glucose), fetal bovine serum (FBS), donor horse serum (DHS), and penicillin-streptomycin (P/S) were purchased from Biological Industries (Israel). Bovine serum albumin (BSA) was obtained from Sigma (USA). Alamar Blue cell proliferation assay was obtained from Thermo Fisher (USA). Immunofluorescence staining was performed by using primary antibodies against myogenin (ab1835) and myosin heavy chain, obtained from Abcam (UK). Secondary antibodies goat anti-mouse (Alexa Fluor^® 488) (ab150113) and goat anti-rabbit (Alexa Fluor^® 594) (ab150080) were purchased from Abcam (UK). Phalloidin (Alexa Fluor^® 488) and DAPI were obtained from Invitrogen (USA).
The Cystoseira algae species used in the preparation of CM-03 were collected from the Marmara Sea (Turkey). MWCNT used for catalysis was purchased from Sigma Aldrich.