Jak2 antibody

Resveratrol was obtained from Macklin (Shanghai, China). DSPE-PEG₂₀₀₀, FA-PEG₂₀₀₀-DSPE was obtained from Ruixibio (Xi’an, China). Soy phosphatidylcholine, cholesterol, DiR iodide were purchased from Meilunbio (Dalian, China). Coumarin 6 (Cou-6) was obtained from Aladdin (Shanghai, China). JAK 2 antibody, P-STAT 3 antibody, Caspase 3 antibody, Cleaved Caspase 3 antibody were purchased from Cell Signaling Technology (Danvers, MA, USA). P-JAK 2 antibody was acquired from Abcam (Cambridge, UK). STAT 3 antibody, BCL-2 antibody, Bax antibody, Cytochrome antibody, and N Tubulin β polyclonal antibody were obtained from Bioworld Technology (Nanjing, China). The peroxidase-conjugated secondary antibody was obtained from CWBIO (Taizhou, China).
Human osteosarcoma cell line 143B cells were obtained from the Nanfang Hospital, Southern Medical University (Guangzhou, China). Cells were cultured in DMEM medium containing 10% FBS in a 37°C incubator containing 5% CO₂.
Male nude mice (6–8 weeks old) were purchased from Bestest (Zhuhai, China). All animal studies were conducted in accordance with the Guide for the Care and Use of Laboratory Animals. The Experimental Animal Ethics Committee of South China Hospital, South China University approved all experimental protocols.

Tumour tissues and cell samples were lysed with protein lysis buffer and centrifuged at 4 °C. Protein lysates were divided by 10% SDS-PAGE and transferred onto a PVDF membrane. The membrane was blocked with 5% skim milk at room temperature, followed by overnight incubation at 4 °C on a shaker with METTL3 antibody (1:1000, Cell Signalling Technology, E3F2A), JAK2 antibody (1:1000, Cell Signalling Technology, D2E12), STAT3 antibody (1:1000, Cell Signalling Technology, D3Z2G), p-STAT3 antibody (1:1000, Cell Signalling Technology, D3A7). After washing with TBST, the membranes were incubated with secondary antibodies at room temperature. SuperSignal West Pico chemiluminescent substrate (Thermo Scientific, 34094) was used for detection. β-actin (1:2000, Cell Signalling Technology, 8H10D10) was applied as an internal reference. Protein bands were scanned and quantified using ImageJ software via densitometry analysis.