Fig 1: (A) Overexpression of human transcription factor 19 (TCF19) in INS-1 cells was confirmed by qRT-PCR. (B) Western blot against anti-myc tag confirms overexpression of human TCF19 (hsTCF19) (C) Overexpression of hsTCF19 in INS-1 cells leads to increased 3H-thymidine incorporation (n = 5). (D) hsTCF19 overexpression does not lead to any significant changes in cell cycle gene expression (n = 5) (E) Overexpression of hsTCF19 in INS-1 cells does not affect cell viability (n = 5). Data are means ± SEM * p < 0.05.
Fig 2: MAGIC analysis on the list of upregulated genes after TCF19 expression in INS-1 cells identifies significant enrichment for genes with known ChIP signals for STAT1, STAT2, and IRF1 in their promoters (FDR < 10%). A higher score reflects increased likelihood that the factor is enriched for binding gene regions in the list of upregulated genes. Genes included in analysis were those that were upregulated more than 2-fold with an associated FDR < 5%.
Fig 3: Overexpression of TCF19 in INS-1 cells does not lead to increased activation of transcription factor targets but leads to decreased activation of STAT1 Y701 phosphorylation. Representative images of two western blot replicates along with analysis of all replicates are shown. Densitometry with Image J 1.44o was used to quantify the bands on the western blots, which were then normalized to the housekeeper protein band. (A) Serine 727 phospho-STAT1/STAT1 protein expression does not show a statistically significant difference between control and hsTCF19 overexpressing cells (n = 5) (phospho-STAT1~91 kDa, STAT1~84, 91 kDa). (B) Tyrosine 701 phospho-STAT1/STAT1 protein expression shows statistically significant decrease in hsTCF19 overexpressing cells compared to control (n = 3) (phospho-STAT1~84, 91 kDa). (C) IRF1 protein levels are not significantly different. Representative western blots in the figure have IRF1 levels normalized to GAPDH. Three other replicates are normalized to beta tubulin (n = 5) (IRF1~48 kDa). (D) There is no difference in phospho NF-κB/NF-κB levels with hsTCF19 overexpression (n = 3) (phospho NF-κB~65 kDa, NF-κB~65 kDa). All data are means ± SEM * p < 0.05.
Fig 4: Human TCF19 overexpression in human islets upregulates key DNA damage repair genes as well as viral response genes. (A) hsTCF19 overexpression in human islets was confirmed with qRT-PCR. Control represents human islets transfected with an empty vector and hsTCF19 represents human islets transfected with TCF19 plasmid. (B) hsTCF19 overexpression in human islets leads to upregulation of genes (qRT-PCR) that were also upregulated in INS-1 cells (n = 5). Gene names are shown on the horizontal axis. Data are means ± SEM * p < 0.05.
Supplier Page from DNASU for TCF19 (Homo sapiens) in pDNR-Dual (Creator donor/master vector)