Supplementary Fig. 6: Lnczc3h7a facilitates RIG-I signaling pathway by promoting the interaction of TRIM25 and RIG-I.
From: The long noncoding RNA Lnczc3h7a promotes a TRIM25-mediated RIG-I antiviral innate immune response
a, Co-immunoprecipitation analysis of oligomerization and K48-linked ubiquitination of TRIM25 in HEK293T cells co-transfected with flag-TRIM25, V5-TRIM25, HA-Ub-K48 and Lnczc3h7a for 24 hr, infected with VSV for 8 hr. IP, immunoprecipitation; WCL, whole cell lysates. b, Co-immunoprecipitation analysis of the interaction of TRIM25 and MAVS and K48-linked ubiquitination of MAVS in HEK 293T cells co-transfected with flag-TRIM25, V5-MAVS, HA-Ub-K48 and Lnczc3h7a for 24 hr, infected with VSV for 8 hr. c, Co-immunoprecipitation analysis of the interaction of TRIM25 with RIG-I and K63-linked ubiquitination of RIG-I in HEK 293T cells co-transfected with flag-RIG-I, V5-TRIM25, HA-Ub-K63 and Lnczc3h7a for 24 hr, infected with VSV for 8 hr. d, Immunofluorescence assay of the co-localization of endogenous K63-Ub (green) and RIG-I (red) in Lnczc3h7a+/+ and Lnczc3h7a-/- mice peritoneal macrophages upon VSV infection for 8 hr. Nuclei were stained with DAPI (blue). Scale bar, 5 μm. e, Co-immunoprecipitation analysis of the interaction of RIG-I and MAVS in the presence of TRIM25 or Lnczc3h7a in HEK 293T cells co-transfected with flag-RIG-I, V5-MAVS, Myc-TRIM25 and Lnczc3h7a for 24 hr, and infected with VSV for 8 hr. f. Schematic illustration of the location of Trim25 sgRNA target sequence (top) and frameshift mutation sequence (below). Data are representative of three independent experiments (a-e).
