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Western blotting analysis Tissues or cells were resuspended in lysis buffer on ice for 5 minutes, and the supernatants were collected after centrifugation at 13,000 g for 15 minutes

sgRNA constructs and gene expressing codon optimized (OPT) plasmid constructs sgRNA control and constructs against CBS and L2HGDH were designed targeting GTATTACTGATATTGGTGGG (sgControl) (#230080, Addgene), GGAGCAGACAACCTACGAGG (sg CBS -1) (#230081, Addgene), TGGCAGTGCTGGCAGCACGG (sg CBS -2) (#230082, Addgene), GATATAGTCATCGTTGGTGG (sg L2HGDH -1) (#230083, Addgene), and CACCAGACTGGACATAACAG (sg L2HGDH -2) (#230084, Addgene) and constructed in lentiCRISPR v2-Blast (#83480, Addgene) for Blasticidin selection at a final concentration of 5 g/ml

Water suppression was achieved using the VAPOR sequence (Tk et al., 1999)

Mutations in genes such as SNCA, LRRK2, PARK7, PINK1, and VPS35 are associated with familial forms of PD, while pesticide exposure, heavy metals, and industrial chemicals contribute to sporadic cases