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. 2023 Oct 25;15(11):2147. doi: 10.3390/v15112147

Figure 3.

Figure 3

Pteropus vampyrus TRIM40 interacts with RIG-I-like receptors RIG-I and MDA5. Cells were co-transfected with Pteropus vampyrus HA-PV-TRIM40 and FLAG-PV-RIG-I-like receptor (RLR) RIG-I or MDA5 for 30 h in 293T (A) or ZFBK13-75 cells (B). The whole cell extracts (WCE) were used to check levels of expression and immunoprecipitated (IP) with anti-FLAG antibody beads. (C) 293T cells were transfected with human (Hu) or PV HA-TRIM40 and partially purified using HA peptide. Separate 293T cells were transfected with Hu- or PV-RLRs or HA-Hu-IRF3. Lysates from FLAG-protein expressing cells were immunoprecipitated with FLAG beads and washed prior to the addition of HA-purified TRIM40. (D) ZFBK13-75 cells were co-transfected with FLAG-PV-RIG-I and HA-PV-TRIM40 for 24 h and then stimulated with 1 μg/mL low-molecular weight (LMW) poly(I:C) for 1 or 4 h, and lysates were used for WCE or IP: FLAG. (E) ZFBK13-75 cells were co-transfected with FLAG-PV-MDA5 and HA-PV-TRIM40 for 24 h and then stimulated with 1 ug/mL high-molecular weight (HMW) poly(I:C) for 1 or 4 h, and lysates were used for WCE or IP: FLAG.