Ddk tag

Commercially available antibodies were used to detect FK2 (Enzo Life Sciences, BML-PW8810), DDK tag (Origene, TA50011), HCV NS3 (TORDJI-22, Abcam), HCV core (ThermoFisher Scientific, MA1-080), HAV capsid (Commonwealth Serum Laboratories, K24F2), Dengue capsid (Dr. Tom Hobman, University of Alberta), STAT1 (Cell Signaling Technologies, CST9175), STAT2 (Santa Cruz, sc-476), Y701-phosphoSTAT1 (Cell Signaling Technologies, 9167S), Y690-phosphoSTAT2 (Cell Signaling Technologies, 88410S), PDLIM2 (Abcam, 246868), p65 NF-κB (Santa Cruz, sc-372), Actin (EMD Millipore, MAB1501), PARP-1 (BD Pharmingen, BD556362), Lamin (Zymed, 33–2000), B-tubulin (Abcam, AB6046), Anti-NS5a (gift from Charlie Rice, 9E10). For western blotting, goat anti-mouse IR Dye 800 (Licor, 926–32210) and goat anti-rabbit IR Dye 680 (Licor, 926–32221) were used. For immunofluorescence, Alexa Fluor 546 goat anti-mouse IgG (ThermoFisher Scientific, A11030), Alexa Fluor 488 goat anti-rabbit IgG, (ThermoFisher Scientific, A11008), or Alexa Fluor 647 goat anti-mouse IgG (ThermoFisher Scientific, A21236) were used.

Antibodies were from the following sources: N-KDM3A (12835; Proteintech), C-KDM3A (NB100-77282; Novus Biologicals), anti–acetylated α-tubulin (T7451; Sigma-Aldrich), γ-tubulin (GTU-88; Sigma-Aldrich), IFT88 (13967-1-AP; Proteintech), IFT81 (11744-1-AP; Proteintech), GAPDH (5019A-2; ImGENEX), ARL13B (17711-1-AP; Proteintech), DDK tag (TA50011; OriGene), and Halo (G9281; Promega). Secondary antibodies for immunofluorescence and immunoblotting are Fab′2 IgG Alexa Fluor from Molecular Probes and HRP conjugated from EMD Millipore and Sigma-Aldrich.

Western blot was performed as previously described^{18 (link)}. Briefly, half of the left lobe of mouse lungs were homogenized in 300 μl of lysis buffer (1× Reporter lysis buffer (Promega, Madison, WI, USA), containing protease inhibitor (Roche, Basel, Switzerland)) using a TissueLyser II (Qiagen, Germantown, MD, USA). Supernatants were used for SDS-PAGE and western blots. Thirty micrograms of total protein were loaded on 10% SDS-PAGE gels, transferred onto PVDF membrane and probed with primary antibodies against DDK tag (Origene, Rockville, MD, USA), ZO-1, occludin, and VE-cadherin (Invitrogen, Rockford, IL, USA), β1-Na⁺, K⁺-ATPase (Millipore, Billerica, MA, USA), MRCKα (Santa Cruz, Dallas, TX, USA) and glyceraldehyde-3-phosphate dehydrogenase GAPDH (EMD Millipore, Burlington, MA, USA). Data were analyzed using the NIH Image J software.