Plko 3g gfp shrna plasmids

We purchased the pLKO.3G GFP-shRNA plasmids from Addgene. After 48 h of transfection, the viruses were collected from the medium. Cells were infected with the collected viruses over 2 days with the company of polybrene, which were then sorted via GFP signals for 4 days. The specific shRNA sequences were listed as follows: KAT2A-shRNA#1 Sense: 5-GATCCGTACCGGAGACACCAAGCAGGTCTATTTCTCGAG AAATAGACCTGCTTGGTGTCTTTTTTTGG-3; Anti-sense: 5-AATTCCAAAAAAAGACACCAAGCAGGTCTATTTCTCGAG AAATAGACCTGCTTGGTGTCTCCGGTACG-3. KAT2A-shR NA#2 Sense: 5-GATCCCCGGGCTGAACTTTGTGCAGTACAA CTCGAGTTGTACTGCACAAAGTTCAGCTTTTTGG-3; Anti -sense: 5-AATTCCAAAAAGCTGAACTTTGTGCAGTACAAC TCGAGTTGTACTGCACAAAGTTCAGCCCGGG-3. MCT1- shRNA#1 Sense: 5-GATCCCCGGGCTCCGTATTGTTTGA AACATCTCGAGATGTTTCAAACAATACGGAGCTTTTTGG-3; Anti-sense: 5-AATTCCAAAAAGCTCCGTATTGTTTGA  A A C A T CTCGAGATGTTTCAAACAATACGGAGCCCGGG-3; MCT1-shRNA#2 Sense: 5-GATCCCCGGTATGTTTCTGCTAG CTATATACTC G A G T A TATAGCTAGCAGAAACATATTTTTG G-3; Anti-sense: 5-AATTCCAAAAATATGTTTCTGCTAGCTA TATACTCGAGTATATAGCTAGCAGAAACATACCGGG-3.

We purchased the pLKO.3G GFP‐shRNA plasmids from Addgene. After 48 h of transfection, the virus supernatant was obtained through centrifugation. We infected the cells with collected viruses combined with polybrene for 48 h, which were sorted by GFP signals. After seeded in the 24‐well plates, the kidney cancer cell lines (786‐O and ACHN) were then transfected with 20 nM siRNA oligos via RNAiMax reagent (Life Technology). Western blot assays were used to confirm the knockdown efficiency. The specific sequences of siRNA targets are summarized in Table S1.

The pLKO.3G GFP-shRNA plasmids were purchased from Addgene. The shRNA sequence of sh-SPOP#1: 5’-GGAGAACGCUGCAGAAAUU-3’; sh-SPOP#2: 5’-ATAAGTCCAATAACGACAGGC-3’; shINF2-#1: 5’- CCCUCUGUGGUCAACUACU-3’; shINF2-#2 (target to CAAX isoform only): 5’-ACAAAGAAACTGTGTGTGTGA-3’;²³ shDRP1: 5’-GCCAGCUAGAUAUUAACAACAAGAA-3’. shControl: 5’- ACAGACUUCGGAGUACCUG-3’. Viruses were collected from the medium 48 hr after transfection. For knockdown experiments, cells were infected with the collected viruses over 48 hr in the presence of polybrene, followed by GFP sorting for 3–4 days. pTsin- lentivirus vectors were used for overexpression of HA(FLAG)-SPOP-WT or mutants.