Supplementary Figure S3 from Targeting AEBP1 to Mitigate Protumor Activity of Cancer-Associated Fibroblasts and Increase Therapeutic Efficacy of Anti–PD-1

Supplementary figure S3 shows the generation of AEBP1 knockout mouse fibrablasts. (A) Expression levels of long and short isoform transcripts of AEBP1 in NIH3T3 wild-type (WT) fibroblasts.(B) Sanger sequencing results confirming homozygous AEBP1 knockout (KO) clones derived from NIH3T3 mouse fibroblasts.(C) RT-PCR validation of three NIH3T3 AEBP1.KO clones: F5, G5, and H7.(D) CTG cell proliferation assay comparing monocultures of NIH3T3 WT fibroblasts and AEBP1.KO clones F5, G5, and H7 (N = 5 technical replicates). Percentage cell viability was normalized to luminescence data from WT fibroblasts.(E) Venn diagram illustrating significantly down-regulated genes in AEBP1.KO clones F5, G5, and H7 relative to WT cells. Significance thresholds: log2(fold change) < -0.75 and adjusted p-value