RNA Isolation and qRT-PCR Analysis

Total RNA was isolated from tissue samples using the acid guanidium thiocyanate/phenol/chloroform method and was purified with lithium chloride, as previously reported [11 (link),22 (link)]. cDNAs were synthesized using a ImProm Ⅱ Reverse-Transcription system (Promega Corporation, Madison, WI, USA), and real-time polymerase chain reactions were performed on a StepOne Plus Real-time PCR System (Thermo Fisher Scientific, Waltham, MA, USA) using Power SYBR Green PCR Master Mix (Thermo Fisher Scientific) [11 (link)]. Primer sequences for mouse Cdn15 were 5′-TCT TTC TAG GCA TGG TGG GA-3′ and 5′-TCA GTA GTG ATG TTG ACG GC-3′, and those for mouse Il6, Il17a, Tnf, Il1b, and Rn18s (the gene encoding 18S ribosomal RNA) were previously reported [22 (link),54 (link)]. The mRNA values were normalized to the Rn18s levels.