Journal: Nature Communications
Article Title: Epithelial cells release adenosine to promote local TNF production in response to polarity disruption
Figure Lengend Snippet: AdoR promotes transcription of TNF Egr in polarity-deficient epithelial cells. a sal > ent2 , AdoR expression enhances TNF egr transcript levels in the wing disc independently of JNK signalling. TNFR grnd expression, measured by RT-qPCR, is shown as a control. Fold changes are relative to rp49 , n = 6. b – d sal > ent2, AdoR expression upregulates expression of Egr-GFP from a genomic fosmid ( b , c ). This effect is not rescued by co-expression of JNK bskDN ( d ). The arrowhead in b indicates a suspected position effect of the attP site (VK00033) where the fosmid was integrated. e CADO treatment increases expression of TNF egr in explanted wild-type discs, but not in AdoR mutant discs. This effect is further enhanced by Ador overexpression. TNFR grnd expression, measured by RT-qPCR, is shown as a control. Fold changes are relative to rp49 , n ≥ 5. f en > scrib Ri discs upregulate TNF egr mRNA in an AdoR/Ent2-dependent and JNK-independent fashion. Fold changes are relative to rp49 , n ≥ 4. g Human HaCaT cells treated with adenosine upregulates TNF-α transcript levels (as assayed by qRT-PCR). Fold changes are relative to GAPDH , n = 5. Expression of PKG1 , a housekeeping gene and survivin , a JNK target gene, are also shown. h The effect shown in g was suppressed by three different adenosine receptor antagonists (CGS15943, SCH58261 or Caffeine) and by a PKA inhibitor cocktail (Merck # 20-114), but not by JNK inhibitor SP600125. Fold changes are relative to GAPDH , n ≥ 3 and expression of PKG1 is shown. i , j AdoR mutants live longer than wild-type flies when continuously fed with a low dose of Bleomycin ( i ) but live shorter than wild-type flies during acute desiccation (dry starvation, j ). **** P
Article Snippet: Adenosine, SCH58261, Caffeine and the JNK inhibitor SP600125 were obtained from Sigma.
Techniques: Expressing, Quantitative RT-PCR, Mutagenesis, Over Expression