Journal: bioRxiv

Article Title: A local translation program regulates centriole amplification in the airway epithelium

doi: 10.1101/2022.01.18.476821

Figure Lengend Snippet: a) Each panel represents a 3D image of confocal Z-stack images of a single MCC in culture. Apically localized granules are highlighted by dashed lines. TNRC6A stained with human Index serum (18033). DCP1A, DDX6 and XRN1 are concentrated in randomly-localized granules (P-bodies). However, DCP1A and DDX6 are undetectable in apically granules. XRN1 are present in small number of apically granules at low levels. b) Co-localization profile of apically-localized or randomly-localized granules (P-bodies) within the same MCC indicating the different concentration of these proteins in these two classes of granules. Percentage of granules with double labeling signal.

Article Snippet: For IF staining, cryosections or transwell filter of ALI cultures were washed in PBS, and then blocked by 5% donkey serum in PBST (1XPBS, 0.3% Triton X-100) for one hour at RT, followed by incubation with primary antibodies overnight at 4°C (TNRC6A, human index serum, 1:3000; FOXJ1, eBioscience, # 14-9965-82, 1:100; Ac-α-tub, Sigma, # T7451, 1:10,000, or Abcam # ab125356; DCP1A, Abcam, #ab183709, 1:50; DDX6, Cell Signaling Technology, #8988S, 1:20; EDC3, Santa Cruz, # sc-55081, 1:20; EDC4/GE-1, Cell Signaling Technology, #2548S, 1:50; Xrn1, Bethyl Laboratories, #A300-443A, 1:20; γ-tubulin, Abcam, #ab84355, 1:50; EIF3B, Santa Cruz , # sc-16377, 1:50).

Techniques: Staining, Concentration Assay, Labeling

Journal: eLife

Article Title: Fragile X mental retardation protein is a Zika virus restriction factor that is antagonized by subgenomic flaviviral RNA

doi: 10.7554/eLife.39023

Figure Lengend Snippet: ( A ) Schematic illustrating the secondary structures present in the ZIKV 3′ UTR and RNAs used for affinity chromatography. Deletion mutant RNA constructs fused to a tobramycin aptamer at the 5′ end are shown. DENV NS2A coding sequence was used as a negative control RNA. ( B ) Purified RNAs bound to tobramycin-sepharose beads were incubated with HeLa cell lysate and unbound proteins were washed away prior to elution for western blotting for DDX6, FMRP, FXR1, FXR2, G3BP1 and PTB.

Article Snippet: The following antibodies were used for IP, western blotting and/or immunofluorescence analysis: anti-envelope protein 4G2 , rabbit IgG (2729S, Cell Signaling Technologies), anti-FMRP (ab17722, ABCAM, Cambridge, UK), anti-FXR1 (12295S, Cell Signaling Technologies), anti-FXR2 (7098S, Cell Signaling Technologies), anti-G3BP1 (A302-033A, Bethyl Laboratories), rabbit anti-DDX6 (9407S, Cell Signaling technologies), rabbit anti-PTB (homemade), rabbit anti-ZIKV NS4B (GTX133321, Genetex), anti-ZIKV NS2B (GTX133308, Genetex), anti-BRD4 (13440, Cell Signaling Technologies), anti-GAPDH (ab9485, ABCAM), anti-TLN1 (SC-365875, Santa Cruz Biotechnology), anti-PNPLA6 (SC-271049, Santa Cruz Biotechnology).

Techniques: Affinity Chromatography, Mutagenesis, Construct, Sequencing, Negative Control, Purification, Incubation, Western Blot

Journal: eLife

Article Title: Fragile X mental retardation protein is a Zika virus restriction factor that is antagonized by subgenomic flaviviral RNA

doi: 10.7554/eLife.39023

Figure Lengend Snippet:

Article Snippet: The following antibodies were used for IP, western blotting and/or immunofluorescence analysis: anti-envelope protein 4G2 , rabbit IgG (2729S, Cell Signaling Technologies), anti-FMRP (ab17722, ABCAM, Cambridge, UK), anti-FXR1 (12295S, Cell Signaling Technologies), anti-FXR2 (7098S, Cell Signaling Technologies), anti-G3BP1 (A302-033A, Bethyl Laboratories), rabbit anti-DDX6 (9407S, Cell Signaling technologies), rabbit anti-PTB (homemade), rabbit anti-ZIKV NS4B (GTX133321, Genetex), anti-ZIKV NS2B (GTX133308, Genetex), anti-BRD4 (13440, Cell Signaling Technologies), anti-GAPDH (ab9485, ABCAM), anti-TLN1 (SC-365875, Santa Cruz Biotechnology), anti-PNPLA6 (SC-271049, Santa Cruz Biotechnology).

Techniques: Knock-Out, Sequencing, Northern Blot

Journal: eLife

Article Title: Fragile X mental retardation protein is a Zika virus restriction factor that is antagonized by subgenomic flaviviral RNA

doi: 10.7554/eLife.39023

Figure Lengend Snippet: ( A ) Schematic illustrating the secondary structures present in the ZIKV 3′ UTR and RNAs used for affinity chromatography. Deletion mutant RNA constructs fused to a tobramycin aptamer at the 5′ end are shown. DENV NS2A coding sequence was used as a negative control RNA. ( B ) Purified RNAs bound to tobramycin-sepharose beads were incubated with HeLa cell lysate and unbound proteins were washed away prior to elution for western blotting for DDX6, FMRP, FXR1, FXR2, G3BP1 and PTB.

Article Snippet: Antibody , Anti-DDX6 (Rabbit polyclonal) , Cell Signaling Technology , Cell Signaling Technology Cat# 9407S, RRID: AB_10556959 , (1:1000).

Techniques: Affinity Chromatography, Mutagenesis, Construct, Sequencing, Negative Control, Purification, Incubation, Western Blot

Journal: eLife

Article Title: Fragile X mental retardation protein is a Zika virus restriction factor that is antagonized by subgenomic flaviviral RNA

doi: 10.7554/eLife.39023

Figure Lengend Snippet:

Article Snippet: Antibody , Anti-DDX6 (Rabbit polyclonal) , Cell Signaling Technology , Cell Signaling Technology Cat# 9407S, RRID: AB_10556959 , (1:1000).

Techniques: Knock-Out, Sequencing, Northern Blot

Journal: eLife

Article Title: Fragile X mental retardation protein is a Zika virus restriction factor that is antagonized by subgenomic flaviviral RNA

doi: 10.7554/eLife.39023

Figure Lengend Snippet:

Article Snippet: Antibody , Anti-DDX6 (Rabbit polyclonal) , Cell Signaling Technology , Cell Signaling Technology Cat# 9407S, RRID: AB_10556959 , (1:1000).

Techniques: Knock-Out, Sequencing, Northern Blot