Journal:

Article Title: Proneural Gene ash1 Promotes Amacrine Cell Production in the Chick Retina

doi: 10.1002/dneu.20693

Figure Lengend Snippet: Expression of ash1 in the developing chick retina detected with dig-labeled antisense RNA probe. A: ash1 expression in E6 retina. B: ash1 expression at E8. C: Higher magnification of a peripheral region at E8. D: Higher magnification of a central region at E8. E–G: Immunostaining to mark the anatomical locations in a pseudostratified E8 retina of photoreceptor cells (Pr, visinin+, E), amacrine cells (Am, AP2α+, F), and ganglion cells (Gc, Islet-1+, G). H–K: Comparison of the spatial distribution of cells expressing ash1 (H, J) with that of cells expressing ngn2 (I, K), in developmental stage-matched regions of E8 retinas (H, I; and J, K). The neuroepithelial zone (NE) within the developing retina is marked, approximately, by a vertical line. Scale bars: 100 μm.

Article Snippet: Additional antibodies included: rabbit polyclonal antibodies against viral protein p27 (1:1000 or 1:200 for retinal sections already subjected to in situ hybridization; Spafas) and against AP2 α (1:200; Santa Cruz, Biotechnology, Santa Cruz, CA), and monoclonal antibodies against Brn3a (1:200; Chemicon, Temecula, CA) and RA4 (1:500 dilution; a gift from Steven McLoon, University of Minnesota, Minneapolis, MN).

Techniques: Expressing, Labeling, Immunostaining, Comparison

Journal:

Article Title: Proneural Gene ash1 Promotes Amacrine Cell Production in the Chick Retina

doi: 10.1002/dneu.20693

Figure Lengend Snippet: Expansion of amacrine territories in retinas infected with RCAS-ash1. A, B: Immuno-staining for viral protein p27 in E12 retinas thoroughly (A) or partially (B) infected by RCAS-ngn2. C, D: Double-labeling of an E8 retina thoroughly infected with RCAS-ash1 for viral protein p27 (C) and for AP2α (D). E, F: Double-labeling of an E10 control retina partially infected with RCAS-GFP for p27 to identify viral infection (E) and for AP2α to identify amacrine cells (F). G, H: Double-labeling of an E10 retina partially infected with RCAS-ash1 for p27 (G) and for AP2α (H). I, J: Double-labeling of an E10 control retina partially infected with RCAS-GFP for p27 (I) and for Pax6 to identify amacrine cells (J). K, L: Double-labeling of an E10 retina partially infected with RCAS-ash1 for p27 (K) and for Pax6 (L). Purple bars mark the domain thickness of AP2α+ cells (G, H) or Pax6+ cells (K, L). Yellow lines mark virally infected regions. The anatomical location of each major cell type is marked on the right. Pr, photoreceptors; Bi, bipolar cells; Am, amacrine cells; Gc, ganglion cells. Scale bars: 100 μm.

Article Snippet: Additional antibodies included: rabbit polyclonal antibodies against viral protein p27 (1:1000 or 1:200 for retinal sections already subjected to in situ hybridization; Spafas) and against AP2 α (1:200; Santa Cruz, Biotechnology, Santa Cruz, CA), and monoclonal antibodies against Brn3a (1:200; Chemicon, Temecula, CA) and RA4 (1:500 dilution; a gift from Steven McLoon, University of Minnesota, Minneapolis, MN).

Techniques: Infection, Immunostaining, Labeling, Control

Journal:

Article Title: Proneural Gene ash1 Promotes Amacrine Cell Production in the Chick Retina

doi: 10.1002/dneu.20693

Figure Lengend Snippet: Quantitative analyses of the effect of ash1 overexpression on various retinal cell populations. A: A plot of the ratios of immuno-positive cells in infected regions over adjacent, uninfected regions in retinal sections. B: The percentage of cells positive for each marker. Data were obtained using dissociated E9 retinal cells (or E8 for RCAS-ash1ΔCrb) seeded into culture dishes. AP2, AP2α for amacrine cells; Pax6 for amacrine cells; Brn3A for ganglion cells; LIM for horizontal cells; Vis, visinin for photoreceptor cells; Vim, vimentin for Müller glia; chx10 mRNA for bipolar cells; BrdU for proliferating cells; TUNEL for apoptotic cells. Shown are means ± SDs. “*” indicates statistically significant at p < 0.05 level, and “**” at p < 0.01 level.

Article Snippet: Additional antibodies included: rabbit polyclonal antibodies against viral protein p27 (1:1000 or 1:200 for retinal sections already subjected to in situ hybridization; Spafas) and against AP2 α (1:200; Santa Cruz, Biotechnology, Santa Cruz, CA), and monoclonal antibodies against Brn3a (1:200; Chemicon, Temecula, CA) and RA4 (1:500 dilution; a gift from Steven McLoon, University of Minnesota, Minneapolis, MN).

Techniques: Over Expression, Infection, Marker, TUNEL Assay

Journal:

Article Title: Proneural Gene ash1 Promotes Amacrine Cell Production in the Chick Retina

doi: 10.1002/dneu.20693

Figure Lengend Snippet: Lack of amacrine overexpression in retinas infected with RCAS-ngn2 and RCAS-ath5. A, B: Double-labeling of an E10 retina infected with RCAS-ngn2 for viral protein p27 to label infected regions (A) and for AP2α to identify amacrine cells (B). C, D: Immunostaining for calretinin of RPE cell cultures infected with control virus RCAS-GFP (C) or RCAS-ngn2 (D). E–H: Double-labeling of an E10 retina infected with RCAS-ath5 for viral protein p27 (E, G) and for AP2α to label amacrine cells (F) or for Brn3A to identify ganglion cells (H). Infected regions are indicated by lines. The anatomical location of each major cell type is marked on the right. Pr, photoreceptors; Bi, bipolar cells; Am, amacrine cells; Gc, ganglion cells. Scale bars: 100 μm.

Article Snippet: Additional antibodies included: rabbit polyclonal antibodies against viral protein p27 (1:1000 or 1:200 for retinal sections already subjected to in situ hybridization; Spafas) and against AP2 α (1:200; Santa Cruz, Biotechnology, Santa Cruz, CA), and monoclonal antibodies against Brn3a (1:200; Chemicon, Temecula, CA) and RA4 (1:500 dilution; a gift from Steven McLoon, University of Minnesota, Minneapolis, MN).

Techniques: Over Expression, Infection, Labeling, Immunostaining, Control, Virus

Journal:

Article Title: Proneural Gene ash1 Promotes Amacrine Cell Production in the Chick Retina

doi: 10.1002/dneu.20693

Figure Lengend Snippet: Varied effects on embryo development of different constructs derived from ash1. A: A schematic diagram showing the structural regions included in each construct. B: E14 embryos infected with RCAS-ash1ΔC (left) or RCAS-ash1ΔN (right). C: E12 embryos infected with RCAS-ash1ΔCrb (left) or RCAS-GFP (right). D: A comparison of an eye from an E12 embryo infected with RCAS-ash1ΔCrb (left) with the control (right). Arrows point to regions with hypo-pigmentation. E: A plot of the ratios of immuno-positive cells in infected regions over those in the adjacent, uninfected regions. Data were obtained from scoring the number of immuno-positive cells in retinal sections. Cell markers are: AP2 (AP2α) and Pax6 for amacrine; Brn3A for ganglion; LIM for horizontal; Vis (visinin) for photoreceptor; Vim (vimentin) for Müller glia; BrdU for proliferating; and TUNEL for apoptotic cells. Shown are means ± SDs. “*” indicates statistically significant at p < 0.05 level, and “**” at p < 0.01 level.

Article Snippet: Additional antibodies included: rabbit polyclonal antibodies against viral protein p27 (1:1000 or 1:200 for retinal sections already subjected to in situ hybridization; Spafas) and against AP2 α (1:200; Santa Cruz, Biotechnology, Santa Cruz, CA), and monoclonal antibodies against Brn3a (1:200; Chemicon, Temecula, CA) and RA4 (1:500 dilution; a gift from Steven McLoon, University of Minnesota, Minneapolis, MN).

Techniques: Construct, Derivative Assay, Infection, Comparison, Control, TUNEL Assay

Journal:

Article Title: Proneural Gene ash1 Promotes Amacrine Cell Production in the Chick Retina

doi: 10.1002/dneu.20693

Figure Lengend Snippet: Differential effects on retinal cell populations between RCAS-ash1ΔN and RCAS-ash1ΔCrb. A–H: Double-labeling of E8 (A–D) or E10 (E–H) retinas infected with RCAS-ash1ΔN for viral protein p27 to identify infected regions (A, C, E, G) and for BrdU (B), ngn2 expression (D), Pax6 (F), or TUNEL (H). I–O: Double-labeling of E8 (I–L) or 10 retinas (M–O) infected with RCAS-ash1ΔCrb for viral protein p27 to identify infected regions (I, K, M, O) and for BrdU (J), ngn2 expression (L), AP2α (N), or TUNEL (O). Infected regions are marked by lines. Scale bars: 100 μm.

Article Snippet: Additional antibodies included: rabbit polyclonal antibodies against viral protein p27 (1:1000 or 1:200 for retinal sections already subjected to in situ hybridization; Spafas) and against AP2 α (1:200; Santa Cruz, Biotechnology, Santa Cruz, CA), and monoclonal antibodies against Brn3a (1:200; Chemicon, Temecula, CA) and RA4 (1:500 dilution; a gift from Steven McLoon, University of Minnesota, Minneapolis, MN).

Techniques: Labeling, Infection, Expressing, TUNEL Assay

Journal: Microbiology Spectrum

Article Title: Attenuate host susceptibility to respiratory virus invasion by inhibiting interactions between host proteins SLC16A3 and AP1G1

doi: 10.1128/spectrum.03116-24

Figure Lengend Snippet: The interactions between SLC16A3 and AP1G1 ( A ) TPCA-MS experimental scheme. ( B–D ) SLC16A3 and AP1G1 exhibit similar melting curves. Cells were non-infected or infected with H1N1, RSV, and 229E viruses. ( E ) IP-WB result demonstrating the interaction between SLC16A3 and AP1G1. ( F–H ) Immunofluorescence images illustrating the subcellular localization of SLC16A3 and AP1G1. BEAS2B cells were infected with H1N1, RSV, and 229E viruses. ( I–K ) Western blot assay showing the subcellular localization of AP1G1. In = input, C = cytoplasm, M = cytomembrane. Cells were infected with H1N1, RSV, and 229E virus (MOI = 2, n = 3).

Article Snippet: Primary antibodies of AP1G1 (NBP3-15727) were purchased from Novus Biologicals (Shanghai, China), and SLC16A2 (20676-1-AP) was obtained from Proteintech (Wuhan, China).

Techniques: Infection, Immunofluorescence, Western Blot, Virus

Journal: Microbiology Spectrum

Article Title: Attenuate host susceptibility to respiratory virus invasion by inhibiting interactions between host proteins SLC16A3 and AP1G1

doi: 10.1128/spectrum.03116-24

Figure Lengend Snippet: SFJD intervention with SLC16A3-AP1G1 interaction. ( A ) KEGG Strip Pathway. ( B ) Lactate expression levels in BALF of mice before and after SFJD administration after infection with H1N1 virus. t -test, * P < 0.05, ** P < 0.01, n = 6. ( C ) SLC16A3 expression levels in lung tissue of mice before and after SFJD administration after infection with H1N1 virus ( n = 6). ( D ) IP-WB assay illustrating the attenuated interaction between SLC16A3 and AP1G1 following SFJD treatment ( n = 3). ( E–G ) Subcellular localization of AP1G1 before and after SFJD administration following infection with H1N1, RSV, and 229E viruses. Cells were infected with H1N1, RSV, and 229E virus (MOI = 2, n = 3).

Article Snippet: Primary antibodies of AP1G1 (NBP3-15727) were purchased from Novus Biologicals (Shanghai, China), and SLC16A2 (20676-1-AP) was obtained from Proteintech (Wuhan, China).

Techniques: Stripping Membranes, Expressing, Infection, Virus