vsx2  (Proteintech)

Journal: Cell & Bioscience

Article Title: NOTCH2NLC GGC repeat expansions cause retinal neurodegeneration in neuronal intranuclear inclusion disease mouse model

doi: 10.1186/s13578-026-01542-x

Figure Lengend Snippet: Expression of NOTCH2NLC-polyG in the retina of NIID mice. A – F Co-immunostaining on retinas of NIID and control mice using anti-Flag (indicating NOTCH2NLC-polyG) with anti-VSX2 ( A ), anti-PAX6 ( B ), anti-RBPMS ( C ), anti-Rhodopsin ( D ), anti-Arrestin C ( E ), or anti-RPE65 ( F ). Red: VSX2 ( A ), PAX6 ( B ), RBPMS ( C ), Rhodopsin ( D ), Arrestin C ( E ) or RPE65 ( F ); Green: NOTCH2NLC-polyG; Blue: DAPI. Scale bar = 50 μm. The yellow dashed box indicates the region shown at higher magnification. G The percentage of cells co-expressing PolyG and VSX2, PAX6 or RBPMS. H The quantification of the number of VSX2-, PAX6-, and RBPMS-positive cells in NIID and control mice. Data are presented as mean ± SEM. N = 6 per group, ** P = 0.0012 (PAX6), ** P = 0.0021 (RBPMS), ns = no significance, two-tailed t-test. I Transmission electron microscopy images of intranuclear inclusions in the INL and RGC layer of the retinas of NIID mice. The yellow dashed box indicates the region shown at higher magnification. The yellow arrows indicate the round-shaped, filamentous, non-membranous intranuclear inclusions. Scale bar = 1 μm

Article Snippet: Primary antibodies used in this study include: RBPMS (Thermo Fisher Scientific, PA5-31231, 1/300), VSX2 (Proteintech, 25825-1-AP, 1/300), PAX6 (Abcam, ab195045, 1/300), Rhodopsin (Proteintech, 30438-1-AP, 1/200), Arrestin C (Proteintech, 85067-1-AP, 1/200), Flag (Sigma, F1804, 1/300), Flag (Thermo Fisher Scientific, PA5-31231, 1/300), Tuj1 (Sigma, MAB5564, 1/300), GFAP (Cell Signaling Technology, 3670S, 1/300), IBA1 (Wako, 019-19741, 1/300), GFP (Thermo Fisher Scientific, MA5-15256, 1/300).

Techniques: Expressing, Immunostaining, Control, Two Tailed Test, Transmission Assay, Electron Microscopy

Journal: Genes & Development

Article Title: DIO3 coordinates photoreceptor development timing and fate stability in human retinal organoids

doi: 10.1101/gad.352924.125

Figure Lengend Snippet: DIO3 is expressed in RPCs in the developing human retina and retinal organoids. ( A ) UMAP from CZ CellxGene Discover of human fetal retina scRNA-seq with major cell types labeled. ( B ) DIO3 mRNA expression in fetal retina is limited to DIO3 + RPCs, overlaid on the unannotated UMAP. ( C ) DIO3 is expressed in VSX2 + cells in a 14 week old fetal retina. ( D–F ) DIO3 expression in human retinal organoids at days 80 ( D ), 140 ( E ), and 200 ( F ). ( G ) DIO3 and Ki67 expression in human retinal organoids at day 90. Green circles indicate Ki67 + /DIO3 + cells. ( H ) DIO3 and cone opsin expression in human retinal organoids at day 140. Yellow outlines indicate L/M-opsin + cells. Blue outlines indicate S-opsin + cells. ( H ) Region of morphologically naive photoreceptors within an organoid (same organoid as in I ). ( I ) Region of morphologically more mature photoreceptors within an organoid (same organoid as in H ). ( J ) DIO3 activity as measured by deiodination activity of wild-type organoids over time. Error bars represent SEM.

Article Snippet: Antibodies used in this study were as follows: goat ARR3 (1:200; Novus Biologicals NBP1-37003), mouse CRX (1:500; Abnova UJOHO013), mouse CRX (1:200; Sigma-Aldrich WH0001406M2), rabbit DIO3 (1:200; Novus Biologicals NBP1-05767), mouse Ki67 (1:200; Santa Cruz Biotechnology sc-23900), goat NRL (1:200; Bio-Techne AF2945), rabbit OPN1LW/OPN1MW (1:200; Sigma-Aldrich AB5405), chick OPN1LW/OPN1MW (1:200; kind gift from Jeremy Nathans), goat OPN1SW (1:200; Santa Cruz Biotechnology sc-14363), chick OPN1SW (1:200; kind gift from Jeremy Nathans), rabbit RCVRN (1:200; Sigma-Aldrich AB5585), rat RFP (1:400; Proteintech 5F8-150), mouse rhodopsin (1:500; GeneTex GTX23267), mouse rhodopsin (1:500; Thermo Fisher Scientific MA5-11741), and sheep VSX2 (1:100; Exalpha Biologicals X1180P).

Techniques: Labeling, Expressing, Activity Assay