(A) Peritoneal macrophages were isolated from wild-type (WT) mice and infected with vesicular stomatitis virus (VSV) for the indicated hours. Then the Tim-3 mRNA expression was examined by …
Flox gene amplication.
Cre gene amplication.
Genotype identification.
Phenotype identification.
Tim-3 mRNA expression in macrophages in response to VSV infection.
Tim-3 protein expression in macrophage in response to VSV infection.
VSV induces Tim-3 phosphorylation .
Tim-3 signaling promotes VSV replication in macrophage .
(A) Mating protocol. (B and C) Mice tails were cut and DNA was extracted. Flox gene (B) and Cre gene (C) segments were PCR amplified and determined by electrophoresis. (D) Genotype examination to …
(A and B) Protein complex of Tim-3 and NF90 overexpressed in cells. HEK293T cells were transfected with plasmids encoding HA-Tim-3, Flag-NF90, and Myc-Tim-3 for 24 hr, immunoprecipitated with HA or …
Tim-3-WT were transfected into HEK293T cells.
Candidates interacts with Tim-3.
Tim-3 interacts with NF90.
NF90 interacts with Tim-3.
Tim-3 interacts with NF90 via intracellular tail.
Tim-3 inhibits NF90.
(A and B) Co-immunoprecipitation and mass spectrometry for screening of proteins interacting with Tim-3. Tim-3-WT were transfected into HEK293T cells for 24 hr, cell lysates were immunoprecipitated …
(A) Tim-3 enhances the ubiquitination of NF90 in macrophages in response to vesicular stomatitis virus (VSV) challenges. Peritoneal macrophages in wild-type (WT) and Tim-3-TG mice were infected with …
Tim-3 mediates NF90 ubiquitination.
Tim-3 promotes ubiquitination of NF90.
Tim-3 promotes k48-linked ubiquitination of NF90.
NF90 canbe ubiquitinated at the DZF domain.
Tim-3 promotes NF90 ubiquitination at the DZF domain.
The cells were lysed, immunoprecipitated with NF90 (ILF3) antibody, and detected by western blot using the indicated antibodies.
(A) E3 ligases identified by mass spectrometry for top peptide hits (defined by Mascot score) associated with NF90 ubiquitination. (B) TRIM47 promotes NF90 degradation in a proteasome-dependent …
Silence of TRIM47 in macrophages.
TRIM47 promotes NF90 degradation.
TRIM47 promotes NF90 degradation.
Tim-3 synthesizes with TRIM47 to promote NF90 degradation.
Tim-3 promotes TRIM47 expression.
Tim-3 promotes TRIM47 expression.
Tim-3promotes k63-linked ubiquitination of TRIM47.
(A) Quantitative reverse transcription PCR (qPCR) analysis of TRIM47 mRNA expression in RAW264.7 macrophages 48 hr after transfection with TRIM47 siRNA. (B) Western blot analysis of the protein …
(A and B) The intracellular domain of Tim-3 and the DZF domain of NF90 interact with TRIM47 respectively. HEK293T cells were transfected with the indicated plasmids for 24 hr and treated with MG132 …
Lys297 is a critical site for ubiquitination of NF9.
Tim-3 interacts with TRIM47 via its intracellular tail.
NF90 interacts with TRIM47 via its DZF domain.
Tim-3 promotes TRIM47 mediated ubiquitination of NF90.
Lys297 is a critical site for TRIM47-mediated ubiquitination of NF90.
Lys297 is a critical site for TRIM47-mediated ubiquitination of NF90-DZF.
Co-immunoprecipitation analysis of the polyubiquitination of NF90-DZF (WT) and its mutants in HEK293T cells transfected with plasmids encoding HA–ubiquitin (K48). Data are representative of three …
(A) Lysates from wild-type (WT) and Tim-3 gene (Havcr2) knockout (Tim-3KO) peritoneal macrophages infected with vesicular stomatitis virus (VSV) were analyzed by immunoblotting for the indicated …
TRIM47 knock down increases TIA-1,G3BP1 expression and decrease VSV expression.
Tim-3 selectively inhibits the phosphorylation of PKR and eIF2a.
Tim-3 increases the expression of SGs markers G3BP1 and TIA-1 in macrophages.
TRIM47 was silenced in RAW264.7 macrophages as in Figure 2—figure supplement 1. (A and B) Quantitative reverse transcription PCR (qPCR) analysis of TIA-1 and G3BP1 expression in RAW264.7 macrophages …
(A, B, D, E, G, and H) Detection of mRNA transcription of G3BP1 and TIA-1 in organs and peritoneal macrophages by quantitative reverse transcription PCR (qPCR) after wild-type (WT) and Tim-3KO mice …
Tim-3 knowdown increases G3BP1expression in spleen with VSV infection.
Tim-3 knowdown increases TIA-1 expression in spleen with VSV infection.
Tim-3 knowdown decreases VSV expression in spleen with infection.
Tim-3 knowdown increases G3BP1 expression in lung with VSV infection.
Tim-3 knowdown increases TIA-1 expression in lung with VSV infection.
Tim-3 knowdown decreases VSV expression in lung with VSV infection.
Tim-3 knowdown increases G3BP1 expression in peritoneal macrophages with VSV infection.
Tim-3 knowdown increases TIA-1 expression in peritoneal macrophages with VSV infection.
Tim-3 knowdown decreases VSV expression in peritoneal macrophages with VSV infection.
Tim-3 deficiency protects mice from VSV infection.
Tim-3 knowdown attenuated tissue damage.
Sequences of the primers used for PCR.