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Journal: bioRxiv
Article Title: Loss of MTPAP disrupts mitochondrial RNA processing causing upregulation of type I interferon signalling
doi: 10.64898/2026.05.04.722669
Figure Lengend Snippet: A: (Left) Relative levels of mtDNA expressed as the averaged relative levels of MT-CO2 , MT-ND1 and mtDNA D-loop signals and ( Right ) of mtRNA expressed as the averaged relative expression levels of MT-ATP6/8 , MT-ND4 , MT-CO1 , MT-CYTB and MT-ND6 in lysates of control cells and of MTPAP_KO cells (sg#2) treated for 10 days with ddC. Mean ± SEM; n=3 experiments, each colour is a different experiment, ** indicates p<0.01, in two-way ANOVA with Holm-Sidak multiple comparison test. B-C-D-J-L-N: Western blot analysis showing protein levels of MTPAP and IRF3 ( B ), STING ( C ), cGAS ( D ), MAVS ( J ), MDA5 ( L ), RIG-I ( N ), in lysates of double KO cells (sg#2 for MTPAP_KO) compared to controls. Vinculin or cofilin are used as loading controls. Pictures representative of 3 different experiments. E: mRNA levels of IFNB1 measured by qPCR in cGAS/MTPAP and STING/MTPAP double KO BJ-5ta cells compared to controls 24 h after stimulation with transfected HT-DNA (sg#2 for MTPAP_KO). Mean ± SEM; n=3 experiments, each colour is a different experiment, **** indicates p< 0.0001 in two-way ANOVA with Holm-Sidak multiple comparison test. F-H: ISG score measured by qPCR in BJ-5ta cells double KO for STING/MTPAP ( F ) cGAS/MTPAP ( H ) cells compared to controls (sg#2 for MTPAP_KO). Mean ± SEM; n≥4 experiments, each colour is a different experiment, MTPAP_KO using sg#2; ns indicates non significance for Wilcoxon tests. G-I: IFNB1 mRNA expression measured by qPCR in double KO STING/MTPAP ( G ) cGAS/MTPAP ( I ) cells compared to controls (sg#2 for MTPAP_KO). N=4 experiments, each colour is a different experiment; ns indicates non significance for Wilcoxon tests. K-M: IFNB1 mRNA expression levels in MAVS_KO ( K ) and MDA5_KO ( M ) cells compared to BJ-5ta controls when transfected with poly(I:C). Mean ± SEM; n=3 experiments, ** indicates p<0.01, *** p< 0.001 in two-way ANOVA with Holm-Sidak multiple comparison test. O: IFNB1 mRNA expression levels in RIG-I_KO BJ-5ta cells compared to controls Infected with Sendai Virus. Mean ± SEM; n=5, ** indicates p<0.01 in two-way ANOVA with Holm-Sidak multiple comparison test.
Article Snippet: KO of RIG-I was validated by exposing cells to
Techniques: Expressing, Control, Comparison, Western Blot, Transfection, Infection, Virus