Sybr green 1

Total RNA was extracted from renal tissues and cell lines using TRIzol reagent (Invitrogen, Carlsbad, CA, USA), following the manufacturer's protocol. The total RNA was reverse transcribed into complementary DNA (cDNA) using HiScript II (Vazyme, Shanghai, China). qRT-PCR was performed using SYBR Green I (Vazyme, Shanghai, China) on ABI 7900 system (Applied Biosystems, Carlsbad, CA, USA) and the primers for APOC1 were as follows: forward(F), 5′-AGGACAAGGCTCGGGAACTCAT-3′, and reverse(R), 5′-GATGTCACCCTTCAGGTCCTCA-3′. The primers for β-actin were as follows: (F), 5′-GAAGATCAAGATCATTGCTCCT-3′, and (R), 5′-TACTCCTGCTTGCTGATCCA-3′.

Total RNA was drawn from cell lines and tissues by using TRIzol reagent (Invitrogen, Carlsbad, CA, USA). Complementary DNA (cDNA) was synthesized by using PrimeScript RT Master Mix (TaKaRa, Kyoto, Japan). Reverse transcription polymerase chain reaction (RTPCR) was conducted by using HiScript II (Vazyme, Shanghai, China) in accordance with the manufacturer’s guidelines. Quantitative RT-PCR (qRT-PCR) was performed using SYBR Green I (Vazyme, Shanghai, China) on ABI 7900 system (Applied Biosystems, Carlsbad, CA, USA). β-actin was employed as a normalization. For PCR, we used the following primers (Realgene, Nanjing, China): TRIM37 (5′-ATGAGCGTGTTCGGGAAATTAG-3′ and 5′-TCACTCTTACTACAAGACCGCAA-3′); β-actin (5′-ACTGGAACGGTGAAGGTGAC-3′ and 5′-AGAGAAGTGGGGTGGCTTTT-3′).
The Chromatin Immunoprecipitation Kit (Catalog # 17–371, Millipore) was used to perform ChIP assays, in line with the manufacturer’s protocols. Briefly, cells with stable lentiviral transduction were selected and purified for subsequent antibodies immunoprecipitation, antibodies against TRIM37, H2A and negative control IgG as described above were used in the ChIP assays. ChIP DNA products were amplified with specific primer of TGF-β1 promoter (5′-GGCAGTTGGCGAGAACAGT-3′ and 5′-CTGGGGTCAGCTCTGACAGT-3′).

We performed eQTL analysis using 16 brain tissues from patients with drug-resistant epilepsy to investigate whether identified SNPs regulate BCL11A expression in human brain tissues. Tissue RNA Kit (Omega, United States) and the FastQuant cDNA kit (Tiangen, China) are used for genomic RNA extraction and cDNA reverse transcription, respectively, and the SYBR^®Green I (Vazyme, China) and the ABI QuantStudio 6 Flex^™ (ABI, United States) analyzer are used for Quantitative PCR. We designed the primers with Primer Premier V6.0 (Premier Biosoft Inc., United States). Details on Quantitative PCR premiers are provided in Supplementary Table 2. Functional effect of identified SNPs on BCL11A expression was further verified by data from the Genotype-Tissue Expression project (GTEx).²

To examine if the tagging SNPs associated with epilepsy and drug-resistant epilepsy were also associated with STX1B expression level in drug-resistant epileptic brain tissues, we conducted eQTL analysis using temporal lobe samples from 16 patients (nine males and seven females) with drug-resistant epilepsy. Genomic RNA was extracted using the Tissue RNA Kit (OMEGA, United States), and reverse transcribed into cDNA using the FastQuant cDNA kit (Tiangen, China). Quantitative PCR was conducted using SYBR^®Green I (Vazyme, China) and the ABI QuantStudio six Flex™ (ABI, United States) analyzer. Primers were designed using Primer Premier V6.0 (Premier Biosoft Inc., United States) Relative expression levels were determined using the 2^−ΔΔCT method, and differences in STX1B mRNA expression between genotypes were analyzed by independent sample t-test using Graphpad Prism9.0 (www.graphpad- prism.cn). We also conducted eQTL analysis using results from the BrainCloud database (http://eqtl.brainseq.org) to validate eQTL findings from epileptic brain tissues. Details on qPCR primers are provided in Supplementary Table 2.