CircRNA SOD2 motivates non-small cell lungs cancer advancement with EMT via acting as microRNA-2355-5p’s competing endogenous RNA to mediate calmodulin regulated spectrin associated proteins-2

Circular RNA circ_0004277 Inhibits Acute Myeloid Leukemia Progression Through MicroRNA-134-5p / Single stranded DNA binding protein 2

“…The cells on the bottom side of the membrane were fixed with 95% methanol after 24 hours and stained for 15 minutes with 0.5% crystal violet solution (Sigma, St. Louis, MO, USA). Finally, the cells were counted under a microscope in three random fields of view [ 16 ]. In the migration experiment, the procedures are the same, except that the Matrigel was not used.…”

“…SSBP2-WT or SSBP2-MUT reporter vector was co-transfected with miR-134-5p mimic or control miRNA into U937 and KG-1a cells. Ultimately, the relative luciferase activity of each group was measured by a Dual-Luciferase Assay System (Promega, Madison, WI, USA) [ 16 ].…”

“…The magnetic beads were rinsed, and then the complex was incubated with Proteinase K to eliminate proteins. Ultimately, the RNA was isolated from the immunoprecipitate, and qRT-PCR was performed [ 16 ].…”

“…The beads were rinsed twice with pre-chilled lysis buffer, three times with low salt buffer, and once with high salt buffer after being incubated at 4°C for 3 hours. Subsequently, the immunoprecipitated RNA was isolated and purified, and finally qRT-PCR was conducted to detect the enrichment of circ_0004277 [ 16 ].…”

“…Then, the membranes were blocked with 5% defatted milk, and washed with tris buffered saline tween, and then incubated with anti-SSBP2 antibody (1:1,000; Thermo Fisher Scientific, Waltham, MA, USA) at 4°C overnight, then incubated with HRP-conjugated anti-rabbit antibody (1:20,000; Thermo Fisher Scientific, Waltham, MA, USA) at room temperature for 1 h. The BeyoECL Plus kit (Beyotime, Shanghai, China) was used to develop the protein bands. Image J software was applied to determine the gray values of the protein bands [ 16 ].…”