Sulforhodamine b sodium salt srb

Cells were resuspended and seeded in 96-well plate with a density of 1000 cells per well. After the cells adherence, they were fixed with 10% trichloroacetic acid for at least 24 h, after which they were stained by Sulforhodamine B sodium salt (SRB) (Sigma, USA) for 30 min and washed three times by 1% (vol/vol) acetic acid. About 24 hours later after the plates were dry, 150 ul 10 mmol/L Tris were added to each well and the absorbance was measured at 510 nm³¹ .
For EDU staining, cells were seeded at 50–70% confluence and then stained with EDU at 37 °C for 2 hours. After fixed with 4% paraformaldehyde 15 min at room temperature, they were permeabilized with 0.3% triton X-100. The cells were subsequently added with click reaction buffer following with Hoechst staining for 10 min according to protocol (Beyotime, Shanghai, China).
Cell cycle analysis was performed with PI/RNase Staining Buffer (Becton, Dickinson, Franklin Lakes, NJ, USA) and detected by MUSE Cell Analyzer (Merck Millipore, Darmstadt, Germany) according to the manufacturer’s instructions. All experiment were tripled and the present data were presented mean ± SEM.

The lyophilized microalgal biomass of Nannochloropsis gaditana (batch L3250520) was purchased from Cianoalgae SI (Gipuzkoa, Spain). Enzymatic solutions (Viscozyme^® L, Celluclast^® 1.5 L, Saczyme^® Yield) were kindly donated by Novozymes A/S (Bagsvaerd, Denmark). The main components of the enzymatic preparations and their activities are reported in Table 2.
Chloroform, methanol, and ethanol were purchased from Fisher Scientific GmbH (Wien, Austria). Sodium hydrogen carbonate, potassium hydroxide, Triton X, and dimethylsulfoxide (DMSO) were purchased from Carl Roth (Karlsruhe, Germany). Cell culture media and respective supplements were obtained from Gibco Thermo Fisher Scientific (Waltham, MA, USA) and Szabo Scandic (Vienna, Austria). CellTiter-Blue (CTB) 10× concentrate was purchased from Promega (Waldorf, Germany). Sulforhodamine B sodium salt (SRB) and 2′,7′-dichlorofluorescin diacetate (DCFH-DA) were purchased from Sigma-Aldrich (München, Germany). Invitrogen CyQuant LDH Cytotoxicity Assay Kit was bought from Thermo Fisher Scientific (Waltham, MA, USA). Fatty acid methyl esters standard (Supelco 37 FAME Mix) was from Supelco (Bellefonte, PA, USA).

Cells were seeded in 96-well plates at a density of 1500 cells per well. After at least 6 hours, the first dish was fixed with 10% cold trichloroacetic acid for at least 24 hours, and the other plates were fixed every 24 hours. After washing and drying, the plates were stained with Sulforhodamine B sodium salt (SRB) (Sigma, USA) for 20 minutes and washed with 1% (vol/vol) acetic acid. After drying, 150 µL of 10 mmol/L Tris was added and the absorbance was measured using the microplate reader (Thermo Fisher, USA) at 562 nm. The results were analyzed with GraphPad Prism 8.0.2.

Rabbit anti-phospho-ERK1/2 (#4370), rabbit anti-ERK1/2 (#4695), rabbit anti-cyclin D1 (#2978), rabbit anti-phospho-p90RSK (#12032S), rabbit anti-RSK1 (#8408S), rabbit anti-phospho-Akt ser473 (#9271), rabbit anti-Akt (#9272), rabbit anti-vimentin (#5741), and rabbit α-SMA (#19245) antibodies were purchased from Cell Signaling Technology, Inc. (Danvers, MA, USA). Rabbit anti-MMP2 (#AB9015) and rabbit anti-MMP9 (#AB19016) antibodies were purchased from Merck Millipore (Burlington, MA, USA). Mouse anti-PCNA (#sc-56), mouse anti-OPN (#sc-73631), and mouse anti-KLF4 (#sc-393462) antibodies were purchased from Santa Cruz Biotechnology Inc. (Dallas, TX, USA). Rabbit anti-GAPDH (#LF-PA0018) was purchased from AbFrontier (Seoul, Korea). Mouse anti-α-tubulin antibody, crystal violet (#C0775), 2′,7′-dichlorodihydrofluorescein diacetate (DCF-DA), sulforhodamine B sodium salt (SRB; #S1402), trichloroacetic acid (TCA; #T6399), Mito-TEMPO (MT; #SML0737), and Rh1 were purchased from Sigma-Aldrich (St. Louis, MO, USA). Goat anti-rabbit IgG second antibody (#31460) and goat anti-mouse IgG second antibody (#31430) were obtained from Invitrogen (Carlsbad, CA, USA). Phosphate-buffered saline (PBS; #EBA-1105) was purchased from Elpisbio (Daejeon, South Korea). Diphenyleneiodonium chloride (DPI; #0504) and apocynin (APO; #4663) were obtained from Tocris Bio-science (Ellisville, MO, USA).