🧫 Experiment Protocol
Exploratorymotoneuron degenerationmultiple autophagy genesneuronal cell linesproposed
This experiment used RT-qPCR to analyze the expression of multiple autophagy-related genes following trehalose treatment. The study examined TFEB target genes including Ppargc1a, lysosomal hydrolases and membrane proteins (Ctsb, Gla, Lamp2a, Mcoln1, Tpp1), and autophagy-related components (Becn1, Atg10, Atg12, Sqstm1/p62, Map1lc3b, Hspb8, and Bag3). The researchers demonstrated that most of these genes were upregulated in a PPP3- and TFEB-dependent manner, confirming the transcriptional regulation of autophagy by trehalose through the TFEB pathway.
PRIMARY OUTCOME
mRNA expression levels of autophagy genes
EXPECTED OUTCOMES
Significant upregulation of TFEB target genes and autophagy-related genes following trehalose treatment
SUCCESS CRITERIA
Statistically significant increase in mRNA levels of autophagy genes that is dependent on PPP3 and TFEB
PROTOCOL
RNA extraction, reverse transcription, real-time quantitative PCR analysis of multiple autophagy-related genes