Nr3c1

Western blot was performed to assess SMAD4 expression in miR-19b-3p inhibitor and negative control transfected cells and tumors induced in SCID mice. SMAD4 was detected with anti-SMAD4 (Abcam, Cambridge, UK) at a 1:5000 dilution. The level of SMAD4 was normalized to the level of β-actin protein, which was detected by using anti-β-actin (Abcam, Cambridge, UK) at a 1:2000 dilution. Then horseradish peroxidase (HRP)-tagged anti-rabbit or anti-mouse immunoglobulin (Abgent, San Diego, CA, USA), at a dilution of 1:2000, was used to detect the primary antibody. Enhanced chemiluminescence reagent (Merck Millipore, Temecula, CA, USA) was applied to reveal the protein bands. The Image J software (National Institutes of Health, Bethesda, MD, USA) was used to quantify the band intensity. To further explore the effects of the interactions between miR-19b-3p and several mRNA at the protein level, a series of western blot analyses was performed following the aforementioned experimental protocols. Primary antibodies included PRKACB, ATM, CREB3L2, EGLN3, JUN, NR3C1, WEE1, RASSF1 and TGFBR2 (Abcam, Cambridge, UK).

The following primary antibodies were used in this study: STAT1 (Cell Signaling Technology Inc., #14994), β-actin (Abcam, Inc., #ab115777), TRIM25 (#abs143488), Cathepsin S (#abs137723), NR3C1 (#abs100327), CDKN1B (#abs100463), Arginase 1 (#abs127734), ISG15 (#abs113675), and NOX2 (cat. #abs124860), which were from Absin Bioscience.