RIPK1 D147A, N148A, N169A, N186A, L112A, C256A, R258A, L293A, and E295A mutants were generated using the Q5 site-directed mutagenesis kit (New England Biolabs) according to the manufacturer’s instructions. Primers were designed using NEBaseChanger (New England Biolabs). Immunofluoresce...
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Subsequent library preparation has been performed with NEBNext Ultra Directional RNA Library Prep Kit for Illumina (New England Biolabs) according to the manufacturer’s instructions. Quality and quantity were assessed at all steps via capillary electrophoresis (TapeStation, Agilent Technologies) and ...
Transfections and generation of stable transfectants MDCK or MDCK KO cells stably expressing exogenous proteins (GCaMP6S or GFP-MRTF-A) were generated by transfection using Lipofectamine 2000 according to the manufacturer's instructions. 48 h later, single cells express- ing GFP were FACS ...
Total RNA was extracted from 100 mg of ground leaves using the RNeasy Plant Kit (QIAGEN France S.A.S, Courtaboeuf, France) following the manufacturer’s instructions. The quantity of the RNA was determined using a Qbit 4 Fluorometer (Thermo Scientific, Wilmington, NC, USA) and the RNA ...
following DNAse treatment using Invitrogen™ DNaseI (Thermo Fisher Scientific, Darmstadt, Germany) and subsequent first strand cDNA synthesis using Thermo Scientific™ RevertAid Reverse Transcriptase (Thermo Fisher Scientific, Darmstadt, Germany) were performed according to the manufacturer’s instructions....
Cells were then processed for genomic DNA and RNA using the AllPrep DNA/RNA Micro kit accord- ing to manufacturer instructions (Qiagen, 80284). To prepare the pro- tein lysate from the mouse RPE tissue, the dissected mouse posterior eyecup was transferred to a microcentrifuge tube containing ...
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Return of Partnership Income, including recent updates, related forms and instructions on how to file. Form 1065 is used ... Form 1065 (2022) - IRS Complete Schedule K-3 (Form 1065), Part XIII, for each foreign partner subject to section 864(c)(8) on a transfer or distribution. 27....