Journal: bioRxiv

Article Title: N 6 -methyladenosine (m 6 A) and reader protein YTHDF2 enhance innate immune response by mediating DUSP1 mRNA degradation and activating mitogen-activated protein kinases during bacterial and viral infections

doi: 10.1101/2022.12.01.518805

Figure Lengend Snippet: DUSP1 regulates p38 and JNK phosphorylation and expression of innate immune response genes during bacterial and viral infections. (A) DUSP1 knockdown enhanced the p38 and JNK phosphorylation during infection of P. aeruginosa , C. diphtheriae , or HSV-1 or HSV-1 ICP34.5 mutant virus. (B) DUSP1 knockdown enhanced the expression of IL1β, CSF3, TGM2 and SRC genes during infection of P. aeruginosa, C. diphtheriae , or HSV-1 or HSV-1 ICP34.5 mutant virus. (C) DUSP1 knockdown enhanced the protein level of IL1β during infection of P. aeruginosa, C. diphtheriae or HSV-1 ICP34.5 mutant virus.

Article Snippet: The membrane was blocked with 5% milk and then incubated with primary antibody to GAPDH (5174s, CST), p38 (8690S, CST), p-p38 (4511S, CST), ERK (4695S, CST), p-ERK (4370S, CST), JNK (9252S, CST), p-JNK (4668S, CST), DUSP1 (NBP2-67909, Novus), IL1β (AB-401-NA, R&D system), or ALKBH5 (HPA007196, Sigma) overnight at 4 °C.

Techniques: Phospho-proteomics, Expressing, Knockdown, Infection, Mutagenesis, Virus

Journal: bioRxiv

Article Title: N 6 -methyladenosine (m 6 A) and reader protein YTHDF2 enhance innate immune response by mediating DUSP1 mRNA degradation and activating mitogen-activated protein kinases during bacterial and viral infections

doi: 10.1101/2022.12.01.518805

Figure Lengend Snippet: ALKBH5 regulates p38 and JNK phosphorylation and transcription of innate immune response genes during bacterial and viral infections. (A-D) ALKBH5 knockdown enhanced the p38 and JNK phosphorylation during infection of C. diphtheriae (A), P. aeruginosa (B), or HSV-1 (C) or HSV-1 ICP34.5 mutant virus (D). (E) De novo transcription of IL1β, CSF3, TGM2 and SRC genes following ALKBH5 knockdown at 2 hpi of P. aeruginosa examined by nuclear run-on assay. Cells treated with 4-thiouridine for 1 h after ALKBH5 knockdown were infected P. aeruginosa for 4 h and collected for nuclear run-on assay.

Article Snippet: The membrane was blocked with 5% milk and then incubated with primary antibody to GAPDH (5174s, CST), p38 (8690S, CST), p-p38 (4511S, CST), ERK (4695S, CST), p-ERK (4370S, CST), JNK (9252S, CST), p-JNK (4668S, CST), DUSP1 (NBP2-67909, Novus), IL1β (AB-401-NA, R&D system), or ALKBH5 (HPA007196, Sigma) overnight at 4 °C.

Techniques: Phospho-proteomics, Knockdown, Infection, Mutagenesis, Virus, Nuclear Run-on Assay

Journal: bioRxiv

Article Title: N 6 -methyladenosine (m 6 A) and reader protein YTHDF2 enhance innate immune response by mediating DUSP1 mRNA degradation and activating mitogen-activated protein kinases during bacterial and viral infections

doi: 10.1101/2022.12.01.518805

Figure Lengend Snippet: ALKBH5 regulates the expression of innate immune response genes during bacterial and viral infections. (A) ALKBH5 knockdown enhanced the expression levels of IL1β, CSF3, TGM2 and SRC transcripts during P. aeruginosa, C. diphtheriae , or HSV-1 or HSV-1 ICP34.5 mutant virus infection. (B–D) ALKBH5 knockdown enhanced the protein level of IL1β during infection by C. diphtheriae (B), P. aeruginosa (C), or HSV-1 ICP34.5 mutant virus (D). (E) ALKBH5 overexpression inhibited the expression of IL1β, CSF3, TGM2 and SRC genes during P. aeruginosa infection measured by RT-qPCR. (F) ALKBH5 overexpression inhibited the protein level of IL1β during P. aeruginosa infection measured by Western-blotting.

Article Snippet: The membrane was blocked with 5% milk and then incubated with primary antibody to GAPDH (5174s, CST), p38 (8690S, CST), p-p38 (4511S, CST), ERK (4695S, CST), p-ERK (4370S, CST), JNK (9252S, CST), p-JNK (4668S, CST), DUSP1 (NBP2-67909, Novus), IL1β (AB-401-NA, R&D system), or ALKBH5 (HPA007196, Sigma) overnight at 4 °C.

Techniques: Expressing, Knockdown, Mutagenesis, Virus, Infection, Over Expression, Quantitative RT-PCR, Western Blot