Fig. 4: Genomic inactivation of TFEB and TFE3 decreases proliferation of TSC2 KO cells and xenografts. | Nature Communications

Fig. 4: Genomic inactivation of TFEB and TFE3 decreases proliferation of TSC2 KO cells and xenografts.

From: An mTORC1-mediated negative feedback loop constrains amino acid-induced FLCN-Rag activation in renal cells with TSC2 loss

Fig. 4: Genomic inactivation of TFEB and TFE3 decreases proliferation of TSC2 KO cells and xenografts.

a Immunoblotting of TSC2 KO control cells and cells with CRISPR-Cas9-mediated genomic inactivation of TFEB, TFE3, and dual inactivation of TFEB and TFE3, for TFEB, TFE3, LAMP1, and mTORC1 substrates (p-4EBP1 (S65) and p-p70 S6K (T389)). Two clones representing two unique guide RNAs targeting TFEB and TFE3 are shown. b In vivo growth and tumor volume of subcutaneous TSC2 KO xenografts in NSG mice with single or combined genomic deletion of TFEB and TFE3. n = 5 independent biological replicates. Graphs are presented as mean values ± SEM. Statistical analyses were performed using one-way ANOVA with Dunnett’s test for multiple comparisons. p < 0.0001 (TSC2 KO vs T33-B3-1-11) and p = 0.0112 (TSC2 KO vs T31-B3-3). c Representative TFEB and TFE3 immunohistochemistry in TSC2 KO xenografts in NSG mice with single or combined genomic deletion of TFEB and TFE3 (Scale bar = 100 µm). d Gene Set Enrichment Analysis (GSEA) comparing TSC2 KO versus WT xenografts (top panel), TSC2/TFE3 KO xenografts vs TSC2 KO xenografts (2nd panel), TSC2/TFEB KO xenografts vs TSC2 KO xenografts (3rd panel), and TSC2/TFEB/TFE3 KO xenografts vs TSC2 KO xenografts (4th panel), for TFEB/TFE3 transcriptional targets31,40,41,42. Source data are provided as a Source Data file.

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