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. Author manuscript; available in PMC: 2019 Sep 4.
Published in final edited form as: Nat Microbiol. 2019 Mar 4;4(6):985–995. doi: 10.1038/s41564-019-0375-z

Fig. 5. PYM1 is antiviral against flaviviruses and interacts with flavivirus capsid proteins.

Fig. 5

a, Immunoprecipitations in HEK293 cells expressing the indicated constructs (WNV capsid-2X strep, PYM1-V5) were performed with Strep-Tactin resin (IBA) and blotted with the indicated antibodies. Results are representative of two biologically independent experiments. b, Immunoprecipitation of PYM1 from WNV-infected U2OS cells show a specific interaction between PYM1 with WNV capsid, but not with another WNV protein, NS1. Mouse IgG light chain is detected below PYM1 in the PYM1-IP fraction and is indicated by (*). Results are representative of two biologically independent experiments. c, Relative WNV RNA (WNV/18S) following infection upon siRNA depletion of PYM1 with two independent siRNAs (siPYM1–1, siPYM1–2). Infections are normalized to cells treated with siControl (siCON). Shown is the mean±SE, n = three biologically independent experiments. Significance was calculated using an unpaired, two-tailed student’s t-test and is indicated by * p < 0.05. d, Control (siCON) or PYM1-depleted (siPYM1-1/2) U2OS cells were infected with WNV Kunjin. TCID50 values were measured from cell supernatants. Shown is the mean±SE, n = three biologically independent experiments. Significance was calculated using an unpaired, one-tailed student’s t-test and is indicated by * p < 0.05. e, Immunoprecipitations in HEK293 cells expressing the indicated constructs (DENV or ZIKV capsid-2X strep, PYM1-V5) were performed with Strep-Tactin resin (IBA) and blotted with the indicated antibodies. Results are representative of two biologically independent experiments. f, Relative viral RNA (DENV/18S or ZIKV/18S) following infection upon siRNA depletion of PYM1 with two independent siRNAs (siPYM1–1, siPYM1–2). Infections are normalized to cells treated with siControl (siCON). Shown is the mean±SE, n = three biologically independent experiments. Significance was calculated using an unpaired, two-tailed student’s t-test and is indicated by * p < 0.05, ** p < 0.005.