Journal: Journal of Virology

Article Title: Human Papillomavirus Major Capsid Protein L1 Remains Associated with the Incoming Viral Genome throughout the Entry Process

doi: 10.1128/JVI.00537-17

Figure Lengend Snippet: L1 protein associated with condensed chromosomes retains the reactivity of the H16.V5 epitope. (A and B) At 24 hpi, HaCaT cells infected with EdU-labeled pseudovirus were fixed, permeabilized, and treated with or without Click-iT reaction buffer without dye. Next, the cells were incubated with L1 conformationally dependent mouse MAb H16.V5 antibody (green) and AF488-conjugated anti-α-tubulin (white). Lastly, the cells were treated with Click-iT reaction buffer with AF555 dye (red) to stain the EdU-labeled pseudogenome and mounted in DAPI (blue). Note that colocalization of the EdU puncta and L1 protein is denoted as a yellow color.

Article Snippet: The microtubule network was detected using mouse MAb anti-α-tubulin conjugated to Alexa Fluor 488 (AF488; catalog no. 8058S; Cell Signaling).

Techniques: Infection, Labeling, Incubation, Staining

Journal: Journal of Virology

Article Title: Human Papillomavirus Major Capsid Protein L1 Remains Associated with the Incoming Viral Genome throughout the Entry Process

doi: 10.1128/JVI.00537-17

Figure Lengend Snippet: L1 protein associated with condensed chromosomes retains the reactivity of the H16.56E epitope. (A and B) At 24 hpi, HaCaT cells infected with EdU-labeled pseudovirus were fixed, permeabilized, and treated with or without Click-iT reaction buffer without dye. Next, the cells were incubated with L1 conformationally dependent mouse MAb H16.56E antibody (green) and AF488-conjugated anti-α-tubulin (white). Lastly, the cells were treated with Click-iT reaction buffer with AF555 dye (red) to stain the EdU-labeled pseudogenome and mounted in DAPI (blue). Note that colocalization of the EdU puncta and L1 protein is denoted as a yellow color.

Article Snippet: The microtubule network was detected using mouse MAb anti-α-tubulin conjugated to Alexa Fluor 488 (AF488; catalog no. 8058S; Cell Signaling).

Techniques: Infection, Labeling, Incubation, Staining

Journal: Journal of Virology

Article Title: Human Papillomavirus Major Capsid Protein L1 Remains Associated with the Incoming Viral Genome throughout the Entry Process

doi: 10.1128/JVI.00537-17

Figure Lengend Snippet: The linear epitope of the 33L1-7 antibody is hidden in the L1 protein associated with condensed chromosomes. (A and B) At 24 hpi, HaCaT cells infected with EdU-labeled pseudovirus were fixed, permeabilized, and treated with or without with Click-iT reaction buffer without dye. Next, the cells were incubated with mouse MAb 33L1-7 antibody (green) and AF488-conjugated anti-α-tubulin (white). Lastly, the cells were treated with Click-iT reaction buffer with AF555 dye (red) to stain the EdU-labeled pseudogenome and mounted in DAPI (blue). Note that colocalization of the EdU puncta and L1 protein is denoted as a yellow color.

Article Snippet: The microtubule network was detected using mouse MAb anti-α-tubulin conjugated to Alexa Fluor 488 (AF488; catalog no. 8058S; Cell Signaling).

Techniques: Infection, Labeling, Incubation, Staining

Journal: Journal of Virology

Article Title: Human Papillomavirus Major Capsid Protein L1 Remains Associated with the Incoming Viral Genome throughout the Entry Process

doi: 10.1128/JVI.00537-17

Figure Lengend Snippet: Some full-length L1 protein accompanies the viral genome to the nucleus. (A) HeLa cells were infected with HPV16 pseudovirus with or without 1.5 μM Eg5 inhibitor III (Eg5i) and tracked via live-cell imaging using the IncuCyte Zoom for 48 h. Note that the images are depicted at 18 hpi. (B) HaCaT cells were infected with EdU-labeled HPV16 pseudovirus in the presence of 1.5 μM Eg5i. The cells were fixed at 24 hpi and permeabilized with either digitonin at 0.625 μg/ml or 0.5% TX-100 and then treated with AF555 (green) in Click-iT reaction buffer. The cells were permeabilized again with 0.5% TX-100 and treated with AF647 (red) in Click-iT reaction buffer. Lastly, the cells were mounted in DAPI (blue). (C) HaCaT cells were infected with EdU-labeled pseudovirus and treated with 1.5 μM Eg5i. At 24 hpi, the cells were fixed and permeabilized in 0.5% TX-100. Next, the cells were treated with AF555 (red) in Click-iT reaction buffer, followed by incubation with AF488-conjugated anti-α-tubulin (white) and MAb 33L1-7 (green). Lastly, the cells were mounted in DAPI (blue). Note the colocalization between EdU and L1 signal denoted by white arrows. (D) HeLa cells were infected with HPV16 pseudovirus in the presence of 1.5 μM Eg5i. Cells were trypsinized for 2 min, and monoastral cells were collected. Next, the cells were treated with 15 μl of 0.25% trypsin for 1 h at 37°C. The cells were lysed by passage through a 1-ml syringe with a 25-gauge needle 40 times. Cell lysates were incubated for 1 h at 37°C once more, the trypsin was inactivated, and the samples were analyzed by Western blot analysis with a cocktail of HPV16 L1-specific mouse MAbs (IID5, 33L1-7, and 312F).

Article Snippet: The microtubule network was detected using mouse MAb anti-α-tubulin conjugated to Alexa Fluor 488 (AF488; catalog no. 8058S; Cell Signaling).

Techniques: Infection, Live Cell Imaging, Labeling, Incubation, Western Blot

Journal: Cell death discovery

Article Title: Selective molecular inhibition of the HDAC6 ZnF-UBP binding domain impairs multiple myeloma cell function.

doi: 10.1038/s41420-025-02465-1

Figure Lengend Snippet: Fig. 4 The molecular inhibition of the HDAC6 ZnF-UBP binding domain did not impair the HDAC6 expression or its catalytic activity in RPMI 8226 cells but impaired the aggresome formation. A Expression of HDAC6 and acetylated tubulin (Ac-tub (K40)) in wild-type, HDAC6KO

Article Snippet: The following primary antibodies were used: HDAC6 (Cell Signaling Technology, Danvers, MA, USA, #7558, 1:500), acetylated tubulin (Cell Signaling Technology #3971, 1:1000), GADPH (Merck, Burlington, MA, USA, MAB374, 1:500).

Techniques: Inhibition, Binding Assay, Expressing, Activity Assay