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Table 1 Primer sequences for cloning and mutagenesis of the constructs

From: N-terminal functional domain of Gasdermin A3 regulates mitochondrial homeostasis via mitochondrial targeting

Construct name

Method

Primer sequences (5’ → 3’)

Gsdma3

PCR cloning

forward, AGCGTCGACAGACAATGCCTGTGTTTGAGG;

-pENTR2B

reverse, CCGGAATTCGAAGATAGAGCACAATAAGTAAGTGC

Gsdma3I359N -pENTR2B

mutagenesis

forward, GCTAACTGAAGAACAACTGAAGAATCTAGTAA AATCCTTGGAGA

reverse, TCTCCAAGGATTTTACTAGATTCTTCAGTTGTTCTTCAGTTAGC

Gsdma3M1Btlr

mutagenesis

forward, GCTAACTGAAGAACAACTGAAGAATCTAGTAA AATCCTTGGAGA;

-pENTR2B

reverse, TCTCCAAGGATTTTACTAGA TTCTTCAGTTGTTCTTCAGTTAGC

Gsdma3M1Btlr

PCR cloning

forward, AGCGTCGACAGACAATGCCTGTGTTTGAGG;

-pENTR2B

reverse, GCGGA ATTCCCCTCCAAGGATTTTACTAG

N-Gsdma3

PCR cloning

forward, AGCGAATTCAGACAATGCCTGTGTTTGAGG;

-pCDNA3.1-myc-His(-)A

reverse, AGCGAATTCCGCTGTCATTGCAAATGTACGG

C-Gsdma3

PCR cloning

forward, AGCGAATTCATATGCAAACCTTCCCTAAGATCAGG;

-pCDNA3.1-myc-His(-)A

reverse, CCGGAATTCCAAGATAGAGCACATAAGTAAGTGC

C-Gsdma3Rim3

PCR cloning

forward, AGCGAATTCATATGCAAACCTTCCCTAAGATCAGG;

-pCDNA3.1-myc-His(-)A

reverse, CCGGAATTCCAAGATAGAGCACATAAGTAAGTGC

Gsdma3

PCR cloning

forward, TCCGCGGCCGCACCATGCCTGTGTTTGAGGAT;

-pIRES-hrGFP-1a

reverse, CCGGAATTCCAGATAGAGCACAATAAGTAA

Gsdma3Rim3

PCR cloning

forward, TCCGCGGCCGCACCATGCCTGTGTTTGAGGAT;

-pIRES-hrGFP-1a

reverse, CCGGAATTCCAGATAGAGCACAATAAGTAA

N-Gsdma3

PCR cloning

forward, TCCGCGGCCGCACCATGCCTGTGTTTGAGGAT;

-pIRES-hrGFP-1a

reverse, CCGGAATTCCGCTGTCATTGCAAATGTACGG

C-Gsdma3

PCR cloning

forward, TCCGCGGCCGCACGATGCAAACCTTCCCTAAG;

-pIRES-hrGFP-1a

reverse, CCGGAATTCCAGATAGAGCACAATAAGTAA

C-Gsdma3 (no tag)

PCR cloning

forward, AGCGAATTCATATGCAAACCTTCCCTAAGATCAGG;

-pCDNA3.1-myc-His(-)A

reverse, CCGGAATTCAAAGATAGAGCACATAAGTAAGTGC

C-Gsdma3Rim3 (notag)

PCR cloning

forward, AGCGAATTCATATGCAAACCTTCCCTAAGATCAGG;

-pCDNA3.1-myc-His(-)A

reverse, CCGGAATTCAAAGATAGAGCACATAAGTAAGTGC