Rt pcr assay kit

Curdione was purchased from Shanghai yuanye Bio-Technology Co., Ltd. (Shanghai, China), and Bleomycin was obtained from MedChemExpress LLC (NJ, USA). Antibodies against β-actin, Gapdh, α-SMA, fibronectin, collagen I, p-Jnk, Jnk, p-p38, p38, p-Erk1/2, Erk1/2, p-Smad2, p-Smad3, Smad2/3 were purchased from Cell Signal Technology Inc. (MA, USA). BCA Protein Assay Kit was supplied by Boster Biological Technology co. Ltd. (Wuhan, China). RT-PCR assay kit was obtained from Takara (Dalian, China). Hydroxyproline assay kit was obtained from Nanjing Jiancheng Bioengineering Institute (Nanjing, China). Cell Counting Kit-8 was available from DojindoChemical technology co. LTD (Shanghai, China).

RNA of treated HOS cells was isolated using RNAiso Plus (Cat. No. 9108, Takara, Japan), and mRNA (1 mg) was reverse transcribed using an RT-PCR assay kit (Cat. No. 6110A, RR901A, Takara). The primers used in this study were as follows:
human p21: F: 5′-GCCCGTGAGCGATGGAACTTC-3′, R: 5′-CCTGCCTCCTCCCAACTCATCC-3′; human ATF4: F: 5′-GTCTGCCCGTCCCAAACCTTAC-3′, R: 5′-TCCTGCTCCGCCCTCTTCTTC-3′; human GAPDH: F: 5′-AGGTCGGTGTGAACGGATTTG-3′, R: 5′-TGTAGACCATGTAGTTGAGGTCA-3′. GAPDH was used as a control. SYBR green/fluorescein qPCR Master Mix (Cat. No. RR820A, Takara) was used for the PCR reaction. For RT-PCR amplification, 40 cycles were carried out at 95°C for 30 s, 95°C for 5 s, and 60°C for 30 s, using a CFX96 real-time PCR system (Bio-Rad, USA). The ΔΔCt method was used to evaluate the relative expression levels of genes.

Antibodies: Antibodies against fibronectin and collagen I were purchased from Cell Signaling Technology (MA, USA). Antibodies against Sart1, Arginase 1, β-actin, Gapdh and F4/80 were purchased from Proteintech (Hubei, China). Recombinant IL-4 was purchased from BioLegend (CA, USA). Antibodies against STAT6, p-STAT6 and PPAR-γ were obtained from Cell Signaling Technology (MA, USA).
Lipids: Cholesterol and DSPC were acquired from Sigma-Aldrich, Inc. (St. Louis, MO). mPEG2000-DMG (MW2660) was purchased from NOF Co., Ltd. (Kawasaki Japan). The cationic lipidoid C12-200 was generated through ring opening of epoxides by amine substrates with a previously reported method 21 (link). Clodronate liposomes (anionic) were purchased from FormuMax (CA, USA).
Inhibitors: Inhibitors targeting STAT6 (AS1517499) and PPAR-γ (T0070907) were purchased from MedChemExpress (NJ, USA).
BLM was obtained from Hisun Pharmaceutical Co., Ltd (Zhejiang, China). An RT-PCR assay kit was supplied by Takara (Liaoning, China). A hydroxyproline assay kit was obtained from BioVision (CA, USA). Cell Counting Kit-8 was purchased from Dojindo Chemical technology Co. Ltd. (Shanghai, China).