Journal: bioRxiv
Article Title: A cilia-dependent inflammatory programme links bacterial detection to kidney disease
doi: 10.64898/2026.04.24.720658
Figure Lengend Snippet: (A) qPCR quantification of Ccl2 mRNA in Kif3a i-shRNA MDCK cells treated for 6 hours with TLR-1/2 (Pam3), TRL-2/6 (Pam2), TLR-4 (Lipopolysaccharide, LPS) or TLR-5 agonist (Flagellin, FLA). (B) qPCR quantification of Ccl2 mRNA in luciferase (blue), Kif3a (red) or Ift88 (green) i-shRNA MDCK cells treated for 6 hours by NOD1 (Tri-DAP) or NOD2 agonist (MDP). (C) qPCR quantification of Ccl2 mRNA in doxycycline (DOX) inducible luciferase (blue), Kif3a (red) or Ift88 (green) shRNA MDCK cells treated for 6 hours with 13µM ADPh. (D) qPCR quantification of Ccl2 mRNA in Kif3a i-shRNA MDCK cells treated for 4 hours with 500nM DF-006. (E-F) qPCR quantification of Alpk1 (E) and Tifa (F) mRNA in doxycycline (DOX) inducible luciferase (blue), Kif3a (red) or Ift88 (green) shRNA MDCK cells. (G) qPCR quantification of Ccl2 mRNA in Kif3a i-shRNA MDCK cells treated for 30min with 13µM ADPh in presence of digitonin, and stopped 5 hours and a half later. (H) Percentage of cells with TIFAsome formation in HeLa cells upon ADPh, UPEC heat-killed supernatant stimulation from WT (snUTI89 WT) or lacking Hlde (sn UTI89 ΔHlde), an essential enzyme to generate ADPh. (I-J) qPCR quantification of Ccl2 mRNA expression (expressed in percentage of the maximum value) depending on treatment duration (0min, 30min, 1h, 2h, 4h or 6h) after ADPh (I) and UPEC (J) stimulation of Kif3a i-shRNA MDCK cells. (K) qPCR quantification of Ccl2 mRNA in Kif3a i-shRNA MDCK cells treated for 6 hours with UPEC heat-killed supernatant lacking Hlde (sn UTI89 ΔHlde). (A-B, E-F) Ratio paired t test: *P<0.05, **P<0.01, ***P<0.001, ****P<0.00001. Each dot represents an experimental n. AU: arbitrary units. (D, G-H, K) Paired one-way RM ANOVA (log-normal) with Geisser Greenhouse correction followed by Tukey test: *P<0.05, **P<0.01, ***P<0.001, ****P<0.00001. Each dot represents an experimental n. AU: arbitrary units.
Article Snippet: Pam2 (tlrl-pm2s-1, InvivoGen) and Pam3 (tlrl-pms, InvivoGen) were dissolved in endotoxin-free water at 5 mg/ml and added to the cell culture medium at 5μg/mL final concentration during 1 hour of pre-treatment before stimulation.
Techniques: shRNA, Luciferase, Expressing