Genepix4000b chip scanner

Total RNA of the ESCC tissues and adjacent normal tissues were extracted with RNA enzyme inhibitor RNAiso Plus and delivered to Shanghai Kangcheng Biological Co., LTD for chip service. First, rRNA and DNA were removed from 1 μg of total RNA, then the template was amplified and reversely transcribed into fluorescent cDNA with random primers. The labeled cDNA was hybridized with Human LncRNA Array v3.0 (8 × 60K; Arraystar, Rockville, MD, USA), the chip was washed, and the fluorescence signal intensity of the chip was scanned by a GenePix4000B chip scanner (Molecular Devices, San Jose, CA, USA). Finally, GenePix Pro V6.0 (Molecular Devices) was used for hierarchical clustering analysis of raw data. The screening conditions for differentially expressed genes were as follows: |Fold| ≥ 2, p < 0.05, and false discovery rate (FDR) < 0.05.

In the NP-1 and NS groups, the injured sciatic nerves of three model mice were used for each time point at 1, 3, 5, and 7 days after modeling. The protein chip (QAM-CYT-1; Shanghai Huaying Biomedical Technology Co., Ltd., Shanghai, China) was tested after the material was collected to observe the changes of inflammatory factors (IL-1α, -1β, -6, -10, -12, and TNF-α) in the groups.
Protein was extracted from mouse nerve tissue using a Raybiotech kit (RayBiotech, Norcross, GA, USA) according to the manufacturer's instructions. The protein concentration was quantified using the bicinchoninic acid method and then subjected to quality chip detection. After blocking, sample diluent was added. After washing, diluted Biotin-Antibody Cocktail and sample diluent were added and incubated at room temperature. After cleaning and drying, the chip was scanned using a GenePix 4000B chip scanner (Molecular Devices, Sunnyvale, CA, USA).