Skip to main content
. 2020 Oct 8;3:556. doi: 10.1038/s42003-020-01285-6

Fig. 6. CHIKV nsP1 enhances MSR1-ATG12 interaction.

Fig. 6

a Co-immunoprecipitation (co-IP) of FLAG-Atg12 with Myc-Msr1 (mouse) proteins in the presence of nsP1 in HEK293T cells using a mouse monoclonal anti-FLAG antibody, followed by immunoblotting (IB) with a rabbit anti-FLAG or Msr1 antibody. WCE: whole-cell extract. b Co-IP of FLAG-Atg12 with Myc-nsP1 in the presence of Myc-Msr1 in HEK293T cells using a mouse monoclonal anti-FLAG antibody, followed by IB with a rabbit anti-FLAG, nsP1 or Msr1 antibody. c co-IP of FLAG-Atg12 with endogenous ns1 in trophoblasts infected with CHIKV at a multiplicity of infection (MOI) of 0.5 for 12 h using a mouse anti-FLAG antibody (IP), followed by IB with a rabbit anti-FLAG and anti-nsP1 antibody. d Immunofluorescence staining for endogenous CHIKV nsP1 and LC3 in trophoblasts infected with CHIKV at a MOI of 0.5 for 12 h. The cells were counterstained for nuclei by DAPI (blue). The arrows indicate co-localizations. e Immunoblots of Myc-tagged CHIKV proteins after rapamycin (Rapa) treatment. Myc-tagged CHIKV gene expression plasmids were transfected into HEK293T cells for 24 h. The cells were then treated with 100 nM rapamycin with/without 40 μM of chloroquine (CQ) for 24 h. f The immunoblots show P62 knockout by CRISPR-Cas9 in human trophoblasts. g Immunoblots of Myc-tagged CHIKV proteins in trophoblasts after CHIKV infection. Myc-Cap, nsP1 expression plasmids were transfected into trophoblasts for 24 h. The cells were then infected with CHIKV at a MOI of 0.5 for 12 h. The uncropped immunoblots for all figures can be found in Supplementary Fig. 3.