Author: Saul, Vera Vivian; Seibert, Markus; Krüger, Marcus; Jeratsch, Sylvia; Kracht, Michael; Schmitz, Michael Lienhard
Title: ULK1/2 Restricts the Formation of Inducible SINT-Speckles, Membraneless Organelles Controlling the Threshold of TBK1 Activation Document date: 2019_8_6
ID: 5xk3z4ck_62
Snippet: Reconstituted U2OS cells stably expressing Flag-SINTBAD were treated with 1 µM ULK1 and ULK2-specific Accell siRNAs (Dharmacon #A-005049-13-0005 and #A-005396-14-0005) or with a scrambled control RNA (Seibert et al., 2019) according to the protocol using Accell siRNA Delivery Media (Dharmacon #B-005000-500). Three days after siRNA transfection, half of the cells were seeded on coverslips and used for immunofluorescence staining and the remaining.....
Document: Reconstituted U2OS cells stably expressing Flag-SINTBAD were treated with 1 µM ULK1 and ULK2-specific Accell siRNAs (Dharmacon #A-005049-13-0005 and #A-005396-14-0005) or with a scrambled control RNA (Seibert et al., 2019) according to the protocol using Accell siRNA Delivery Media (Dharmacon #B-005000-500). Three days after siRNA transfection, half of the cells were seeded on coverslips and used for immunofluorescence staining and the remaining cells were analyzed for efficient knockdown by real time qPCR, using the following primers specific for ACTB (5´-CATGTACGTTGCTATCCAGGC-3´, 5´-CTCCTTAATGTCACGCACGAT-3´), ULK1 (5´-ACCCCATTACTGCGAACCTGGA-3´, 5´-GCACGAACAGCAGCGTGAAGC-3´) and ULK2 (5´-TCTGCATCACCATGTGCAAGAA-3´, 5´-AACATCTCATCCAGGGCT-3´).
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