Ab185957

Histological sections were produced with 3-mm thickness containing representative samples of endothelium extracted from the thrombus of patients with CTEPH, which was previously stored at -80°C. Transplanted lungs were used as control tissue by using pieces of the pulmonary artery of the donor. All the samples underwent immunohistochemical analysis for the identification of cytoplasmic HSP70 (Abcam, anti-Hsp70 antibody (3A3), #ab5439) and annexin 2 (Abcam, anti-annexin A2 antibody C-terminal, #ab185957), according to the manufacturer’s protocol. Histomorphometric quantification of the expression of the HSP70 and annexin A2 markers and total area quantification randomly of 5 different fields was performed by using a microscope-coupled image analyzer. The system consists of an Olympus-5 camera coupled to an Olympus microscope, from which the images are viewed on a monitor and evaluated on a digital imaging system (Software Image Pro-Plus 6.0).

The primary antibodies used in this experiment were listed as follows: INSM2 (ab91568, Abcam), MYCN (9405 S, CST), ANXA2 (ab185957, Abcam), FASN (10624-2-AP, Proteintech), ACC (3676 S, CST), ACSS (3658 S, CST), SCD (2794 S, CST), SREBP1 (ab3259, Abcam), mTOR (2983 S, CST), Phospho-mTOR (5536 S, CST) and GAPDH (AP0063, Bioworld Technology Inc.).