mrt67307 402  (MedChemExpress)

Journal: Scientific Reports

Article Title: Pseudomonas aeruginosa -derived DnaJ functions as a novel immunomodulator inducing IFNβ via CME–SGK1–IRF3 axis in macrophages

doi: 10.1038/s41598-025-31281-x

Figure Lengend Snippet: DnaJ-induced IFNβ expression is mediated via the TRIF-dependent pathway. ( A , B ) dTHP-1 cells were transfected with siTRIF (150 nM in B ) for 48 h. Knockdown efficiency is shown in the right panel of ( A ). ( C – E ) Cells were pretreated for 1 h with TBK1 IH (MRT67307; 200 nM in E ). Following transfection or pretreatment, cells were treated with DnaJ (1 µg/ml) for 4 h ( A , C , D ) or for the indicated durations ( B , E ). ( F , G ) Cells were treated with DnaJ (1 µg/ml) for 4 h ( F ) or for the indicated durations ( G ). IFNβ mRNA and protein levels were analyzed by qPCR and ELISA, respectively; protein expression was assessed by immunoblotting. The samples used for immunoblotting were obtained from the same experiment, and the original blots/gels are shown in the Supplementary Figure. Data in ( A , C , D , F ) are expressed as means ± SD ( n = 3). Immunoblots are representative of three experiments. *, p < 0.05; **, p < 0.01; ***, p < 0.001 vs. DnaJ treatment alone ( A , C , D ) or CON ( F ).

Article Snippet: T6167923 (MyD88 inhibitor), MRT67307 (TBK1 inhibitor), and MK-2206 (AKT inhibitor) were purchased from MedChemExpress (Monmouth Junction, NJ, USA).

Techniques: Expressing, Transfection, Knockdown, Enzyme-linked Immunosorbent Assay, Western Blot

Journal: Nature Cell Biology

Article Title: DNA fragmentation factor B suppresses interferon to enable cancer persister cell regrowth

doi: 10.1038/s41556-025-01810-x

Figure Lengend Snippet: a , Single-cell RNA-seq Hallmarks IFNα response gene set signature scores in A375 wild-type DFFB and DFFB-KO parental cells and drug-treated cells at the persister and DTEP timepoints. P values were calculated with the two-sided Mann–Whitney test. b , c , Single-cell RNA-seq UMAP of similar numbers of A375 wild-type and DFFB-KO cells at 9 weeks of drug treatment ( b ) with the enriched Hallmarks IFNα response gene set highlighted in red ( c ). d , e , Measurement of the fraction of cells which regrew into DTEP colonies during treatment: A375 DFFB-KO cells treated with 250 nM dabrafenib and 25 nM trametinib for 2 weeks to derive persister cells, then 1 μM JAK inhibitor (JAKi) ruxolitinib was added and all three drugs were maintained for five more weeks ( d ), and PC9 DFFB-KO cells were treated with 300 nM osimertinib and 5 μM ruxolitinib for 5 weeks ( e ). f , Total STAT1 levels in A375 persister cells cotreated for 14 days with 1 μM TBK1 inhibitor MRT67307 or 1 μM JAKi. g , A375 caspase 9-depleted (pooled KO) and DFFA-CR-expressing parental and persister cells analysed for STAT1. h , PC9 wild-type and DFFB-KO persister cells treated with 500 nM osimertinib analysed for STAT1. i , The A375 DFFB-KO cells treated for 7 days with 250 nM dabrafenib and 25 nM trametinib with and without 1.5 μM of IFNα receptor 1 neutralizing antibody (IFNAR1-nAb) and analysed for total STAT1 expression. j , k , A375 wild-type and STAT1-overexpressing cells ( j ) treated with 250 nM dabrafenib and 25 nM trametinib and analysed for DTEP colony formation ( k ). l , m , PC9 wild-type and STAT1-overexpressing cells ( l ) treated with 300 nM osimertinib and analysed for DTEP colony formation ( m ). In d , e , k and m , n = 3 biological replicates, the mean ± s.d. is shown, and the P values were calculated with the two-tailed Student’s t -test.

Article Snippet: JAK inhibitor ruxolitinib (HY-50856), TBK1 inhibitor MRT67307 (HY-13018), thapsigargin (HY-13433), ABT-737 (HY-50907), S63845 (HY-100741) and T-5224 (HY-12270) were purchased from MedChemExpress.

Techniques: RNA Sequencing, MANN-WHITNEY, Expressing, Two Tailed Test

Journal: bioRxiv

Article Title: Multiple orthoflaviviruses secrete sfRNA in mosquito saliva to promote transmission by inhibiting MDA5-mediated early interferon response

doi: 10.1101/2025.01.21.634113

Figure Lengend Snippet: a Pools of saliva were collected, categorized according to sfRNA:gRNA ratio and used to inoculate cells or human skin explants. b-c WNV infection in Huh7 cells at 24 (b) and 48 hpi (c) with low (L), moderate (M) and high (H) concentration of salivary sfRNA. N per category, 2. d Correlation between infection intensity (i.e., WNV gRNA fold change) in Huh7 and sfRNA concentration (i.e., sfRNA:gRNA ratio) in the corresponding saliva used for infection. e WNV infection in skin explants at 24 h post injection with low and high salivary sfRNA concentration. f Correlation between infection intensity in skin explants and sfRNA concentration in the corresponding saliva used for infection. g Cells and skin explants were transfected and injected, respectively, with sfRNA or Control RNA (Ctl.) prior WNV infection. h-j WNV infection in Huh7 (h), HFF1 (i) and U937 (j) cells at 24 and 48 hpi with WNV post sfRNA transfection. k WNV infection in skin explants at 24 hpi with WNV post sfRNA injection. l Mice were intradermally co-injected with WNV and sfRNA or Ctl. RNA. SfRNA alone was injected as control. Skin biopsies were collected at day 1 and clinical signs and survival were recorded daily. N, 14. m WNV infection in mouse skin at 24 h post injection. n-o Clinical signs (n) and survival (o) for mice co-injected with WNV and either sfRNA or Ctl. RNA. b, c, e Bars show mean ± s.e.m. h-j, k, m Bars show geometric mean ± 95% C.I. n Lines show mean ± s.e.m. Dots indicate repeats. Different letters show significant differences and *, p < 0.05; **, p < 0.01 according to post hoc Fisher’s LSD test, T-test or general linear mixed model. h-j, Different letters indicate statistical differences according to Fisher’s LDS test.

Article Snippet: Media of 2 x 10 5 Huh7 cells was supplemented with 10 µM of MRT67307 (MedChemExpress) diluted in water for 1h, then during transfection of monophosphorylated-folded-sfRNA or the control RNA, and infection with WNV as described above.

Techniques: Infection, Concentration Assay, Injection, Transfection, Control

Journal: bioRxiv

Article Title: Multiple orthoflaviviruses secrete sfRNA in mosquito saliva to promote transmission by inhibiting MDA5-mediated early interferon response

doi: 10.1101/2025.01.21.634113

Figure Lengend Snippet: a-d Expression of IFN-β (a), CXCL10 (b), MX1 (c), and IFI6 (d) in Huh7 cells at 24 and 48 h post infection (hpi) with WNV-infected Culex saliva containing different concentrations of sfRNA. Category of sfRNA concentration; L, low; M, moderate; and H, high. N per category, 2. The dotted line indicates the mock-infected values. e-h Expression of IFN-β (e), CXCL10 (f), MX1 (g), and IFI6 (h) in Huh7 cells at 24 and 48 hpi with WNV post sfRNA transfection. i-l Expression of IFN-β (i), CXCL10 (j), MX1 (k), and IFI6 (l) in HFF1 cells at 24 and 48 hpi with WNV post sfRNA transfection. m-p Expression of IFN-β (m), CXCL10 (n), MX1 (o), and IFI6 (p) in U937 cells at 24 and 48 hpi with WNV post sfRNA transfection. q-t Expression of IFN-β (q), CXCL10 (r), MX1 (s), and IFI6 (t) in skin explants at 24 hpi with WNV post sfRNA injection. Ctl., RNA control. Bars show mean ± sem. Repeats are represented by dots. Different letters show significant differences according to post hoc Fisher’s LSD test or T-test.

Article Snippet: Media of 2 x 10 5 Huh7 cells was supplemented with 10 µM of MRT67307 (MedChemExpress) diluted in water for 1h, then during transfection of monophosphorylated-folded-sfRNA or the control RNA, and infection with WNV as described above.

Techniques: Expressing, Infection, Concentration Assay, Transfection, Injection, Control

Journal: bioRxiv

Article Title: Multiple orthoflaviviruses secrete sfRNA in mosquito saliva to promote transmission by inhibiting MDA5-mediated early interferon response

doi: 10.1101/2025.01.21.634113

Figure Lengend Snippet: a Scheme of RIG-I and MDA5 mediated antiviral interferon response. The methods used to disrupt the antiviral response are written in red. b-d WNV gRNA fold change post sfRNA transfection at 24 and 48 hpi in Huh7 cells treated with MRT67037 (b), in RIG-I-deficient Huh7.5 cells (c), and in Huh7.5 cells upon MDA5 silencing (d). Ctl., RNA control. Bars show geometric mean ± 95% C.I. Repeats are indicated by dots. Different letters show significant differences according to post hoc Fisher’s LSD test.

Article Snippet: Media of 2 x 10 5 Huh7 cells was supplemented with 10 µM of MRT67307 (MedChemExpress) diluted in water for 1h, then during transfection of monophosphorylated-folded-sfRNA or the control RNA, and infection with WNV as described above.

Techniques: Transfection, Control