Dapi staining kit

LuCl₃, cesium acetate (CsAc), lactic acid (LA), oleic acid (OA), 1-octadecene (ODE) and Rhodamine B (RhB) were purchased from Macklin. NH₄F, methanol, ethanol and cyclohexane were purchased from Sinopharm Chemical Reagent Co., Ltd. Phosphate buffered solution (PBS), dulbecco’s modified eagle medium (DMEM) and fetal bovine serum (FBS) were obtained from Adamas Life. Cell counting kit-8 (CCK-8), histone H2AX rabbit polyclonal antibody, fluorescein isothiocyanate (FITC), FITC-labeled goat anti-rabbit IgG (H + L), DAPI staining kit, Ki67 staining kit, 2',7'-bis-(2-carboxyethyl)-5-(and-6)-carboxyfluorescein (BCECF) assay kit, 2,7-Dichlorodi-hydrofluorescein diacetate (DCFH-DA) assay kit, ATP assay kit, hematoxylin and eosin (H&E) staining kit, TUNEL apoptosis assay kit, calcein/PI cell viability/cytotoxicity assay kit, Annexin V-FITC/PI apoptosis assay kit were bought from Beyotime.

At different growth-stages, a 2 mL sample of cell culture was taken and cell counting was performed using a hemocytometer with a size of 16 × 25 squares. To determine the dry weight of cells (DCW), the cells was centrifuged at 3500 g for 3 min, and the resulting cells were freeze dried. The glucose content of the supernatant was measured using a SBA-40D biosensor.
The endoplasmic reticulum (ER) and nuclear staining were carried out using the ER staining kit (bestbio BB-441164) and DAPI staining kit (beyotime C1006), respectively. Briefly, after culturing the cells at 25 °C for 16 h, 1 ml cells was harvested and centrifuged to remove the medium. The cells were then washed once with PBS and resuspended in 100 μL of PBS, followed by the addition of 10 μL DAPI staining solution and ER-Tracker Red staining working solution. The cells were incubated at 37 °C for 15–30 min. After centrifugation to remove the supernatant, the cells were washed three times with PBS and observed using a laser scanning confocal microscope (Fluo View™ FV1000, Olympus). For confocal imaging, the fluorescence of ER, DAPI, and GFP was excited at 340, 586, and 488 nm, respectively, and emitted at 488, 616, and 507 nm.

Hainan Asia Pharmaceutical Co., Ltd. (Haikou, China) provided PPD, which had a purity of > 95% as determined by high-performance liquid chromatography. A549 cells and H460 cells were obtained from American Type Culture Collection (Manassas, VA, United States) and cultured at 37°C in a humidified atmosphere of 5% CO₂ with RPMI-1640 medium, which was supplemented with 10% fetal bovine serum, 100 U/mL penicillin, and 100 mg/mL streptomycin. Ang II and EX-527 were purchased from Sigma-Aldrich (St. Louis, MO, United States). The BCA protein assay reagent kit and DAPI staining kit were purchased from Beyotime Institute of Biotechnology (Jiangsu, China). The primary antibodies for E-cadherin, vimentin, and SIRT1 were purchased from Abcam (Cambridge, United Kingdom). Slug and ZEB1 were purchased from Cell Signaling Technology (Danvers, MA, United States). GAPDH was purchased from ZSGB Biotechnology Co., Ltd. (Beijing, China). The secondary antibodies were purchased from Beijing Dingguo Changsheng Biotechnology Co., Ltd. (Beijing, China).