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. 2023 Nov 13;24(12):e57528. doi: 10.15252/embr.202357528

Figure 1. RNF144A promotes DNA virus‐ or exogenous cytosolic DNA‐triggered innate immune responses.

Figure 1

  • A, B
    PMA‐THP1 cells (A) or HaCaT keratinocytes (B) were stimulated with HSV‐1 (MOI = 1) for the indicated periods. Afterward, the cells were lysed for immunoblot assays.
  • C
    HaCaT keratinocytes were transfected with the empty vector (Vec) or the RNF144A expressing plasmid for 24 h and then infected with HSV‐1 for 24 h. The titers of HSV‐1 were determined by standard plaque assay.
  • D
    HaCaT keratinocytes were transfected with the empty vector (Vec) or the RNF144A expressing plasmid and then infected with HSV‐1 (MOI = 1) for 8 h. The cells were then lysed for real‐time PCR assays.
  • E
    HEK293T cells were transfected with Flag‐RNF144A and then transfected with control siRNA (SC) or RNF144A‐specific siRNA (A1, A2, and A3). At 24 h after transfection, the cells were lysed for immunoblot assays (top). PMA‐THP1 cells were transfected with control siRNA (SC) or RNF144A‐specific siRNA (A1, A2, and A3). At 24 h after transfection, the cells were infected with HSV‐1 (MOI = 1) for 8 h and then immunoblot assays were performed (bottom).
  • F
    PMA‐THP1 cells were transfected with control siRNA (SC) or RNF144A‐specific siRNA (A2 and A3). At 24 h after transfection, the cells were infected with HSV‐1 (MOI = 1) for 24 h. The titers of HSV‐1 were determined by standard plaque assays.
  • G
    PMA‐THP1 cells were transfected with control siRNA (SC) or RNF144A‐specific siRNA (A2). At 24 h after transfection, the cells were infected with GFP‐HSV‐1 (MOI = 1) for 24 h and then immunofluorescence assays were performed. Nuclei were stained with DAPI. Scale bar, 500 μm.
  • H
    PMA‐THP1 cells were transfected with control siRNA (SC) or RNF144A‐specific siRNA (A2 and A3). At 24 h after transfection, the cells were infected with HSV‐1 (MOI = 1) for 8 h. The cells were then lysed for real‐time PCR assays.
  • I
    PMA‐THP1 cells were transfected with control siRNA (SC) or RNF144A‐specific siRNA (A2 and A3). At 24 h after transfection, the cells were infected with HSV‐1 (MOI = 1) for 24 h. The supernatants were collected and subjected to ELISA assays.
  • J
    PMA‐THP1 cells were transfected with control siRNA (SC) or RNF144A‐specific siRNA (A2) for 24 h and then infected with HSV‐1 for 8 h. The cells were lysed for immunoblot assays.

Data information: Two‐tailed unpaired Student's t‐test, *P < 0.05, **P < 0.01, ***P < 0.001. Data shown are from two (C, I) or at least three independent biological replicates (A, B, D–H, J). In (D, H), each data point represents a technical replicate. In (C, F, I), each data point represents an independent biological replicate. Error bars are presented as mean ± SD.

Source data are available online for this figure.