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Table 2 Material

From: Vascular miR-181b controls tissue factor-dependent thrombogenicity and inflammation in type 2 diabetes

ApplicationMaterials
ELISATF ELISA kit (Sekisui), d-dimer ELISA kit (Diagnostica Stago), and TAT ELISA kit (Siemens Healthcare)
TransfectionNegative control mimic (Dharmacon), 200 nM miR-181b mimic (hsa-miR-181b-5p, HMI0270, Sigma), 200 nM anti-miR-181b (anti-hsa-miR-181b-5p, HSTUD0270, Sigma), 200 nM inhibitor control (Dharmacon), siRNA against PTEN (L-003023-00-0005), (Dharmacon) or a control siRNA (Dharmacon)
Taqman PCRmiR-181b-5p (001098), mouse asTF (custom-made), mouse flTF (Mm00438856_m1), human VCAM1 (Hs01003372), mouse VCAM1 (Mm00449197_m1), human PTEN (Hs01628827_s1), mouse PTEN (Mm00477208_m1), mouse KPNA4 (Mm00434725_m1), human GAPDH (Hs99999905_m1), mouse GAPDH (Mm99999915_g1), U6snRNA (001973), and self-designed FAM-tagged TaqMan® gene expression assays for human flTF and asTF (for details see [44])
SYBR Green PCRHuman GAPDH (forward 5′-AGCCACATCGCTCAGACAC-3′, reverse 5′-GCCCAATACGACCAAATCC-3′), human KPNA4 Forward 5′-CAGGAGATTCTTCCAGCCCTTTGTGT-3′, Reverse 5′-ATTACCATCTGTATTTGTTCATTGCCAGCATC-3′)
Western blotHuman flTF (Sekisui diagnostics, 4501, goat IgG), human asTF (monoclonal rabbit, kindly provided by Prof. Vladimir Bogdanov, University of Cincinnati), human VCAM1 (Cell signaling, 13662S, rabbit IgG), human-KPNA4 (Abcam, ab6039, goat IgG), human PTEN (Cell signaling, 9559, rabbit IgG), human p-NFκB (Cell signaling, 3033, rabbit IgG), human NFκB (Santa Cruz, sc-109, rabbit IgG), human histone H3 (Cell signaling, 9715, rabbit), mouse and human GAPDH (Calbiochem, CB1001, mouse IgG), mouse TF (Sekisui diagnostics, 4515, rabbit IgG), mouse VCAM1 (Cell signaling, 39036, rabbit IgG)
IFAMouse TF (Sekisui diagnostics, 4515, rabbit IgG), mouse VCAM1 (Cell signalling, 39036, rabbit IgG), Goat anti Rabbit (Dianova), DAPI (Invitrogen), F-actin probe, Alexa Fluor 647 Phalloidin (Thermo Fisher, A12379)