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In the second PCR, 3 µL of the first PCR reaction product was used as the template and was combined with 2U of Taq, 1 × buffer, 1.5 mM MgCl 2 , 0.4 mg/mL BSA, 0.2 mM dNTPs, and 5pmols each of Illumina adapted primers 341F (CCTACGGGAGGCAGCAG) and 518R (ATTACCGCGGCTGCTGG) (primers + Illumina adapters/barcode/priming region as described in supplemental materials of Bartram et al., 2011: ~ 80 bp) 44,45,46

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ER stress might promote NASH by aggravating diet-induced steatosis (Lee et al., 2008, Oyadomari et al., 2008, Rutkowski et al., 2008), and ER stress has also been shown to directly activate inflammatory signaling cascades (Lebeaupin et al., 2018, Özcan et al., 2004, Willy et al., 2015) and to promote hepatocyte cell death (Iracheta-Vellve et al., 2016, Olivares and Henkel, 2015)

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