Doxorubicin

Saline was purchased from SiYao Co., Ltd. (Shijiazhuang, China). Doxorubicin was purchased from ApexBio Technology LLC (Houston, TX, USA). Tanshinone IIA was purchased from Shanghai Shidande Standard Technical Service Co., Ltd. (Shanghai, China). Pravastatin was purchased from Sangong Pharmaceutical (Shanghai) Co., Ltd. (Shanghai, China). 4% paraformaldehyde was from Beijing Applygen Technology Inc. DMSO was purchased from Sigma-Aldrich LLC (Shanghai, China). Dexrazoxane was purchased from Liaoning Norvino Pharmaceutical Co., Ltd. (Liaoning, China) MHY1485 was purchased from Selleck (Houston, TX, USA). DMEM, FBS, and 0.1 mol/L sodium cacodylate buffer were purchased from Beijing BioDee Biotechnology Co., Ltd. (Beijing, China). GFP-mRFP-LC3 adenovirus and GFP-TFEB lentiviral vector were purchased from Hanbio Technology Co., Ltd. (Shanghai, China). All other chemicals were purchased from commercial sources and were maintained at the highest purity available.

SNK6 was kindly provided by Dr. Y. K. J (Seoul National University Hospital, Seoul, Korea) and NK92MI were purchased from American Type Culture Collection (Rockville, MD, USA). SNK6 was cultured in RPMI-1640 medium supplemented with 10% heat-inactivated fetal bovine serum (FBS) and 500 U/mL of interleukin-2. NK92MI was cultured in MEM-α medium supplemented with 20% heat-inactivated FBS. Penicillin and streptomycin (Gibco-BRL, Grand Island, NY, USA) were added to the media and cells were incubated in a humidified 5% CO₂ atmosphere. Small interfering RNAs (siRNAs) were purchased from Bioneer (Daejeon, South Korea). Cells were transiently transfected using Lipofectamine RNAiMAX (Invitrogen, Carlsbad, CA, USA). Doxorubicin was purchased from Apexbio (Boston, MA, USA).

Saline was purchased from SiYao Co., Ltd. (Shijiazhuang, China). Doxorubicin was bought from ApexBio Technology LLC (Houston, TX, United States). DHT was purchased from Shanghai Shidande SDHTdard Technical Service Co., Ltd. (Shanghai, China). Rapamycin was purchased from Sangong Pharmaceutical Co., Ltd. (Shanghai, China). 4% paraformaldehyde was from Beijing Applygen Technology Inc. (Beijing, China). Dimethyl sulfoxide (DMSO) was purchased from Sigma-Aldrich LLC (Shanghai, China). PDTC was purchased from Abmole China Branch. MHY1485 was purchased from Selleck (Houston, TX, United States). Dulbecco’s Modification of Eagle’s Medium (DMEM), Fetal Bovine Serum (FBS), Penicillin, streptomycin, 0.1 mol/L sodium cacodylate buffer, sodium carboxymethylcellulose and DAPI were purchased from Beijing BioDee Biotechnology Co., Ltd. (Beijing, China). GFP-TFEB lentiviral vector were bought from Hanbio Technology Co., Ltd. (Shanghai, China). All other chemicals were purchased from commercial sources.

The iMSCs utilized in this study were derived from induced pluripotent stem cells (iPSCs) (Nuwacell, Cat #RC1001, China), following the previously reported methodology [53 (link)]. To characterize the iMSCs, flow cytometry was employed to analyze the presence of typical MSC markers. The iMSCs were cultured in an incubator at 37 °C and 5% CO₂ using MSC basal medium (Dakewe, China) supplemented with EliteGro™-Advanced serum-free supplement (Dakewe, China).
Doxorubicin (APExBIO, USA) was utilized to induce cell senescence in iMSCs. Specifically, iMSCs were cultured in MSC basal medium supplemented with serum-free supplement and 100 nM Doxorubicin for 2 days to obtain senescent iMSCs exhibiting altered morphology. Control samples consisted of iMSCs cultured in MSC basal medium supplemented with a serum-free supplement and PBS for 2 days.
Senolytics (ABT-263, NMN, and metformin; Absin, China) were employed to modulate the senescent phenotype of iMSC. Specifically, iMSCs (passages 12–14) were treated with these three drugs for 2 days each.

Doxorubicin, paclitaxel, docetaxel and 4-hydroperoxy cyclophosphamide were purchased from ApexBio (Houston, TX, USA). Puromycin was purchased from Sigma-Aldrich (Shanghai, China).

Cell lines were treated with media carrying DMSO (BioShop, catalog DMS666) as a control, doxorubicin (1–2 nM; MilliporeSigma, catalog D1515), cisplatin (100 nM; TOCRIS, catalog 2251), and PARPi MK-4827 (300 nM; APExBIO, catalog A3617) alone or in combination (doxorubicin/PARPi). Where indicated, cells were treated with ROS scavengers, mitoTEMPO (10 μM; MilliporeSigma, catalog SML0737) or pH 7.4 buffered NAC (5 mM; MilliporeSigma, catalog A9165).