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zeb2  (OriGene)


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    OriGene zeb2
    a , UMAP showing FABP7 expression in control and AIRIM variant cells in day 15 organoids. b , IF image of day 15 control and mutant (V190G and R72W) organoids of transitioning NE marker <t>ZEB2</t> and committed RG marker BLBP (encoded by FABP7). Note that the mutant exhibits less expression of BLBP and less-organized nuclei (marked by ZEB2). Scale bar, 20 µm. c , Quantification of mean BLBP IF intensity of day 15 control and mutant (V190G) organoids. * P = 0.0463. Unpaired two-tailed t -test with Welch’s correction. WT, n = 7; V190G, n = 8 imaged regions from two independent batches. Error bars show s.e.m. d , Quantification of mean BLBP IF intensity of day 15 control and mutant (R72W) organoids. **** P = 8.65486 × 10 −5 . Unpaired two-tailed t -test with Welch’s correction. WT, n = 8; R72W, n = 8 imaged regions from two independent batches. Error bars show s.e.m. e , IF image of day 15 control and AIRIM V190G , AIRIM R72W and AFG2B I466M/V245E organoids of the progenitor marker Sox2 and the neuronal differentiation marker TUBB3. Scale bar, 20 μm. Also included are IF images of day 30 control and AIRIM R72W organoids stained for the progenitor marker Sox2 and the neuronal differentiation marker TUBB3. Scale bar, 40 μm. Three independent batches were performed. f , Representative whole-mount organoid IF images showing the morphology of neural progenitor cells (PAX6 + ), around apical (ZO1 + ) lumens, revealed by sparse labelling with GFP in control and mutant (V190G) organoids. Day 10 cells are columnar and exhibit typical NE morphology. Day 15 control cells show a thinning of apical processes (yellow arrows), whereas mutant cells still seem to be columnar. Scale bar, 20 μm. g , Quantification of the length of neural progenitor cells in control and mutant (V190G) organoids at days 10, 13 and 15, showing reduced length of the progenitor cells in mutant compared with control organoids. Cells with clear apical and basal labelling were used for quantification. Mann–Whitney U -test, **** P < 0.0001, two-tailed, multiple-comparison corrected. Day 10 control, n = 10 cells; day 10 V190G, n = 8 cells; day 13 control, n = 14 cells; day 13 V190G, n = 16 cells; day 15 control, n = 8 cells; and day 15 V190G, n = 8 cells. P adj = 0.0714, day 10; P adj = 9.177792 × 10 −7 , day 13; P adj = 1.098352 × 10 −7 , day 15. Error bars show s.e.m. h , Representative whole-mount organoid IF images showing the morphology of neural progenitor cells revealed by sparse labelling with GFP in control and mutant (R72W) organoids. Day 10 cells are columnar and exhibit typical NE morphology. Day 15 control cells show a thinning of apical processes, whereas mutant cells do not. Scale bar, 20 μm. i , Quantification of the length of neural progenitor cells in control and AIRIM R72W organoids at days 10 and 15. Cells with clear apical and basal labelling were used for quantification. Mann–Whitney U -test, *** P = 0.0009, two-tailed, day 10 control, n = 12 cells; day 10 V190G, n = 9 cells; day 15 control, n = 11 cells; and day 15 V190G, n = 10 cells. Error bars show s.e.m.
    Zeb2, supplied by OriGene, used in various techniques. Bioz Stars score: 93/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/mouse+anti+zeb2/pmc12339376-410-17-18?v=OriGene
    Average 93 stars, based on 7 article reviews
    zeb2 - by Bioz Stars, 2026-07
    93/100 stars

    Images

    1) Product Images from "A programmed decline in ribosome levels governs human early neurodevelopment"

    Article Title: A programmed decline in ribosome levels governs human early neurodevelopment

    Journal: Nature Cell Biology

    doi: 10.1038/s41556-025-01708-8

    a , UMAP showing FABP7 expression in control and AIRIM variant cells in day 15 organoids. b , IF image of day 15 control and mutant (V190G and R72W) organoids of transitioning NE marker ZEB2 and committed RG marker BLBP (encoded by FABP7). Note that the mutant exhibits less expression of BLBP and less-organized nuclei (marked by ZEB2). Scale bar, 20 µm. c , Quantification of mean BLBP IF intensity of day 15 control and mutant (V190G) organoids. * P = 0.0463. Unpaired two-tailed t -test with Welch’s correction. WT, n = 7; V190G, n = 8 imaged regions from two independent batches. Error bars show s.e.m. d , Quantification of mean BLBP IF intensity of day 15 control and mutant (R72W) organoids. **** P = 8.65486 × 10 −5 . Unpaired two-tailed t -test with Welch’s correction. WT, n = 8; R72W, n = 8 imaged regions from two independent batches. Error bars show s.e.m. e , IF image of day 15 control and AIRIM V190G , AIRIM R72W and AFG2B I466M/V245E organoids of the progenitor marker Sox2 and the neuronal differentiation marker TUBB3. Scale bar, 20 μm. Also included are IF images of day 30 control and AIRIM R72W organoids stained for the progenitor marker Sox2 and the neuronal differentiation marker TUBB3. Scale bar, 40 μm. Three independent batches were performed. f , Representative whole-mount organoid IF images showing the morphology of neural progenitor cells (PAX6 + ), around apical (ZO1 + ) lumens, revealed by sparse labelling with GFP in control and mutant (V190G) organoids. Day 10 cells are columnar and exhibit typical NE morphology. Day 15 control cells show a thinning of apical processes (yellow arrows), whereas mutant cells still seem to be columnar. Scale bar, 20 μm. g , Quantification of the length of neural progenitor cells in control and mutant (V190G) organoids at days 10, 13 and 15, showing reduced length of the progenitor cells in mutant compared with control organoids. Cells with clear apical and basal labelling were used for quantification. Mann–Whitney U -test, **** P < 0.0001, two-tailed, multiple-comparison corrected. Day 10 control, n = 10 cells; day 10 V190G, n = 8 cells; day 13 control, n = 14 cells; day 13 V190G, n = 16 cells; day 15 control, n = 8 cells; and day 15 V190G, n = 8 cells. P adj = 0.0714, day 10; P adj = 9.177792 × 10 −7 , day 13; P adj = 1.098352 × 10 −7 , day 15. Error bars show s.e.m. h , Representative whole-mount organoid IF images showing the morphology of neural progenitor cells revealed by sparse labelling with GFP in control and mutant (R72W) organoids. Day 10 cells are columnar and exhibit typical NE morphology. Day 15 control cells show a thinning of apical processes, whereas mutant cells do not. Scale bar, 20 μm. i , Quantification of the length of neural progenitor cells in control and AIRIM R72W organoids at days 10 and 15. Cells with clear apical and basal labelling were used for quantification. Mann–Whitney U -test, *** P = 0.0009, two-tailed, day 10 control, n = 12 cells; day 10 V190G, n = 9 cells; day 15 control, n = 11 cells; and day 15 V190G, n = 10 cells. Error bars show s.e.m.
    Figure Legend Snippet: a , UMAP showing FABP7 expression in control and AIRIM variant cells in day 15 organoids. b , IF image of day 15 control and mutant (V190G and R72W) organoids of transitioning NE marker ZEB2 and committed RG marker BLBP (encoded by FABP7). Note that the mutant exhibits less expression of BLBP and less-organized nuclei (marked by ZEB2). Scale bar, 20 µm. c , Quantification of mean BLBP IF intensity of day 15 control and mutant (V190G) organoids. * P = 0.0463. Unpaired two-tailed t -test with Welch’s correction. WT, n = 7; V190G, n = 8 imaged regions from two independent batches. Error bars show s.e.m. d , Quantification of mean BLBP IF intensity of day 15 control and mutant (R72W) organoids. **** P = 8.65486 × 10 −5 . Unpaired two-tailed t -test with Welch’s correction. WT, n = 8; R72W, n = 8 imaged regions from two independent batches. Error bars show s.e.m. e , IF image of day 15 control and AIRIM V190G , AIRIM R72W and AFG2B I466M/V245E organoids of the progenitor marker Sox2 and the neuronal differentiation marker TUBB3. Scale bar, 20 μm. Also included are IF images of day 30 control and AIRIM R72W organoids stained for the progenitor marker Sox2 and the neuronal differentiation marker TUBB3. Scale bar, 40 μm. Three independent batches were performed. f , Representative whole-mount organoid IF images showing the morphology of neural progenitor cells (PAX6 + ), around apical (ZO1 + ) lumens, revealed by sparse labelling with GFP in control and mutant (V190G) organoids. Day 10 cells are columnar and exhibit typical NE morphology. Day 15 control cells show a thinning of apical processes (yellow arrows), whereas mutant cells still seem to be columnar. Scale bar, 20 μm. g , Quantification of the length of neural progenitor cells in control and mutant (V190G) organoids at days 10, 13 and 15, showing reduced length of the progenitor cells in mutant compared with control organoids. Cells with clear apical and basal labelling were used for quantification. Mann–Whitney U -test, **** P < 0.0001, two-tailed, multiple-comparison corrected. Day 10 control, n = 10 cells; day 10 V190G, n = 8 cells; day 13 control, n = 14 cells; day 13 V190G, n = 16 cells; day 15 control, n = 8 cells; and day 15 V190G, n = 8 cells. P adj = 0.0714, day 10; P adj = 9.177792 × 10 −7 , day 13; P adj = 1.098352 × 10 −7 , day 15. Error bars show s.e.m. h , Representative whole-mount organoid IF images showing the morphology of neural progenitor cells revealed by sparse labelling with GFP in control and mutant (R72W) organoids. Day 10 cells are columnar and exhibit typical NE morphology. Day 15 control cells show a thinning of apical processes, whereas mutant cells do not. Scale bar, 20 μm. i , Quantification of the length of neural progenitor cells in control and AIRIM R72W organoids at days 10 and 15. Cells with clear apical and basal labelling were used for quantification. Mann–Whitney U -test, *** P = 0.0009, two-tailed, day 10 control, n = 12 cells; day 10 V190G, n = 9 cells; day 15 control, n = 11 cells; and day 15 V190G, n = 10 cells. Error bars show s.e.m.

    Techniques Used: Expressing, Control, Variant Assay, Mutagenesis, Marker, Two Tailed Test, Staining, MANN-WHITNEY, Comparison

    a , Quantification of bright-field images at day 15. V190G TSC1+/− neural tissue is enlarged relative to V190G TSC1+/+; ** P < 0.01; **** P < 0.0001; Dunnett’s multiple comparison, unpaired two-sided t -test with Welch’s correction. WT TSC1+/+, n = 11; V190G TSC1+/+, n = 12; WT TSC1+/−, n = 37; V190G TSC1+/−, n = 35, from two independent batches; P = 1.734489 × 10 −6 (TSC1+/+ WT versus TSC1+/+ V190G); P = 4.751388 × 10 −6 (TSC1+/+ V190G versus TSC1 +/− V190G); and P = 0.9354378 (TSC1+/− WT versus TSC1+/− V190G). Error bars show s.e.m. b , Representative images showing the TUNEL signal in control TSC1+/+, V190G TSC1+/+, control TSC1+/− and V190G TSC1+/− organoids at day 15. Scale bar, 50 μm. c , Quantification of TUNEL signal of individual NE bud. TUNEL counts were normalized to the area of the imaged bud. **** P < 0.0001, unpaired, two-sided t -test with Welch’s correction. WT TSC1+/+, n = 21; V190G TSC1+/+, n = 22; WT TSC1+/−, n = 17; V190G TSC1+/−, n = 20 NE buds from two independent batches; P = 2.310684 × 10 −7 (TSC1+/+ V190G versus TSC1+/− V190G); and P = 0.1644 (TSC1+/− WT versus TSC1 +/− V190G). Error bars show s.e.m. d , IF image of day 15 control (WT TSC1+/−) and mutant (V190G TSC1+/−) organoids of transitioning NE marker ZEB2 and committed RG marker BLBP. Scale bar, 50 μm. Two independent batch were performed. e , IF image of day 15 control (WT TSC1+/−) and mutant (V190G TSC1+/−) organoids of VIM (green) and mitochondrial matrix HSP60 (red). Scale bar, 20 μm. Two independent batch were performed. f , Quantification of size of day 15 mutant (V190G) organoids treated with vehicle (DMSO) or 1 μM PI3Kα activator UCL-TRO-1938. * P < 0.05. Unpaired two-sided t -test with Welch’s correction. WT + DMSO, n = 7; V190G + DMSO, n = 12; V190G + 1938 1 μM, n = 11, organoids from two independent batches; P = 0.037447. Error bars show s.e.m. g , Quantification of size of day 15 mutant (R72W) organoids treated with vehicle (DMSO) or 1 μM or 2 µM PI3Kα activator UCL-TRO-1938. * P = 0.0373, ** P = 0.0015, **** P = 8.67138 × 10 −7 . Dunnett’s multiple comparison, unpaired t -test with Welch’s correction (two-sided). Control + DMSO, n = 7; R72W + DMSO, n = 6; V190G + 1938 1 μM, n = 6; V190G + 1938 2 μM, n = 6 organoids from two independent batches. Error bars show s.e.m. h , Quantification of TUNEL signal of individual NE bud. TUNEL counts were normalized to the area of the imaged bud. *** P < 0.001, Dunnett’s T3 multiple comparisons test (two-sided). WT + DMSO, n = 9; V190G + DMSO, n = 11; WT + 1938, n = 7; V190G + 1938, n = 13 from two independent batches. P = 0.0008 (V190G + DMSO versus V190G + 1938). Error bars show s.e.m. i , Representative IF images of day 15 control and mutant (V190G, 1938 1 μM and 1938 2 μM) organoids of BLBP (red). Scale bar, 20 μm. j , Quantification of mean BLBP IF intensity of day 15 control and mutant (V190G, 1938 1 μM and 1938 2 μM) organoids. ****V190G versus control, P = 1.60703 × 10 −5 ; ****V190G versus 1938 1 μM, P = 1.39136 × 10 −6 ; ****V190G versus 1938 2 μM, P = 2.35979 × 10 −7 . Dunnett’s multiple comparison unpaired t -test with Welch’s correction (two-sided). Control + DMSO, n = 6; V190G + DMSO, n = 6; control + 1938, n = 6; V190G + 1938, n = 6 imaged regions from two independent batches, error bars are s.e.m. k , Representative IF images of day 15 control and mutant (R72W, 1938 1 μM and 1938 2 μM) organoids of BLBP (red). Scale bar, 20 μm. l , Quantification of mean BLBP immunofluorescence intensity of day 15 control and mutant (R72W, 1938 1 μM and 1938 2 μM) organoids. **R72W versus 1938 1 μM, P = 0.0027; **R72W versus 1938 2 μM, P = 0.0037; ****R72W versus control, P = 1.49216 × 10 −6 . Dunnett’s multiple comparison, unpaired two-sided t -test with Welch’s correction. Control + DMSO, n = 6; R72W + DMSO, n = 6; R72W + 1938, n = 5; R72W + 1938, n = 6 imaged regions from two independent batches. Error bars show s.e.m. m , IF images of day 15 control (V190G and DMSO 0.1%) and treated (V190G, UCL-TRO-1938 1 μM) organoids stained for VIM (green) and mitochondrial matrix HSP60 (red). Scale bar, 20 μm. n , Quantification of the nuclear/ cytoplasm ratio of RSL24D1 IF of control + DMSO, AIRIM V190G + DMSO, AIRIM V190G + 1938 2 μM, AIRIM V190G TSC1+/− and control + DMSO, AIRIM R72W + DMSO, AIRIM R72W + 1938 2 μM day 15 cerebral organoids. ****AIRIMV190G versus control, P = 1.26963 × 10 −5 ; * AIRIM V190G versus 1938 2 μM, P = 0.024; ** AIRIM V190G versus AIRIM V190G TSC1 +/−, P = 0.0018; **** AIRIM R72W versus control, P = 1.0138 × 10 −5 ; * AIRIM R72W versus 1938 2 μM, P = 0.0491. Dunnett’s multiple comparison, unpaired two-sided t -test with Welch’s correction. Control (V190G + DMSO), n = 12; V190G + DMSO, n = 2; V190G + 1938 2 μM, n = 6; V190G TSC1+/−, n = 12; control (R72W + DMSO), n = 8; R72W + DMSO, n = 11; V190G + 1938 2 μM, n = 12 organoids from two independent batches. Error bars show s.e.m. o , Model describing a potential explanation for the observed suppression of AIRIM V190G phenotypes by UCL-TRO-1938.
    Figure Legend Snippet: a , Quantification of bright-field images at day 15. V190G TSC1+/− neural tissue is enlarged relative to V190G TSC1+/+; ** P < 0.01; **** P < 0.0001; Dunnett’s multiple comparison, unpaired two-sided t -test with Welch’s correction. WT TSC1+/+, n = 11; V190G TSC1+/+, n = 12; WT TSC1+/−, n = 37; V190G TSC1+/−, n = 35, from two independent batches; P = 1.734489 × 10 −6 (TSC1+/+ WT versus TSC1+/+ V190G); P = 4.751388 × 10 −6 (TSC1+/+ V190G versus TSC1 +/− V190G); and P = 0.9354378 (TSC1+/− WT versus TSC1+/− V190G). Error bars show s.e.m. b , Representative images showing the TUNEL signal in control TSC1+/+, V190G TSC1+/+, control TSC1+/− and V190G TSC1+/− organoids at day 15. Scale bar, 50 μm. c , Quantification of TUNEL signal of individual NE bud. TUNEL counts were normalized to the area of the imaged bud. **** P < 0.0001, unpaired, two-sided t -test with Welch’s correction. WT TSC1+/+, n = 21; V190G TSC1+/+, n = 22; WT TSC1+/−, n = 17; V190G TSC1+/−, n = 20 NE buds from two independent batches; P = 2.310684 × 10 −7 (TSC1+/+ V190G versus TSC1+/− V190G); and P = 0.1644 (TSC1+/− WT versus TSC1 +/− V190G). Error bars show s.e.m. d , IF image of day 15 control (WT TSC1+/−) and mutant (V190G TSC1+/−) organoids of transitioning NE marker ZEB2 and committed RG marker BLBP. Scale bar, 50 μm. Two independent batch were performed. e , IF image of day 15 control (WT TSC1+/−) and mutant (V190G TSC1+/−) organoids of VIM (green) and mitochondrial matrix HSP60 (red). Scale bar, 20 μm. Two independent batch were performed. f , Quantification of size of day 15 mutant (V190G) organoids treated with vehicle (DMSO) or 1 μM PI3Kα activator UCL-TRO-1938. * P < 0.05. Unpaired two-sided t -test with Welch’s correction. WT + DMSO, n = 7; V190G + DMSO, n = 12; V190G + 1938 1 μM, n = 11, organoids from two independent batches; P = 0.037447. Error bars show s.e.m. g , Quantification of size of day 15 mutant (R72W) organoids treated with vehicle (DMSO) or 1 μM or 2 µM PI3Kα activator UCL-TRO-1938. * P = 0.0373, ** P = 0.0015, **** P = 8.67138 × 10 −7 . Dunnett’s multiple comparison, unpaired t -test with Welch’s correction (two-sided). Control + DMSO, n = 7; R72W + DMSO, n = 6; V190G + 1938 1 μM, n = 6; V190G + 1938 2 μM, n = 6 organoids from two independent batches. Error bars show s.e.m. h , Quantification of TUNEL signal of individual NE bud. TUNEL counts were normalized to the area of the imaged bud. *** P < 0.001, Dunnett’s T3 multiple comparisons test (two-sided). WT + DMSO, n = 9; V190G + DMSO, n = 11; WT + 1938, n = 7; V190G + 1938, n = 13 from two independent batches. P = 0.0008 (V190G + DMSO versus V190G + 1938). Error bars show s.e.m. i , Representative IF images of day 15 control and mutant (V190G, 1938 1 μM and 1938 2 μM) organoids of BLBP (red). Scale bar, 20 μm. j , Quantification of mean BLBP IF intensity of day 15 control and mutant (V190G, 1938 1 μM and 1938 2 μM) organoids. ****V190G versus control, P = 1.60703 × 10 −5 ; ****V190G versus 1938 1 μM, P = 1.39136 × 10 −6 ; ****V190G versus 1938 2 μM, P = 2.35979 × 10 −7 . Dunnett’s multiple comparison unpaired t -test with Welch’s correction (two-sided). Control + DMSO, n = 6; V190G + DMSO, n = 6; control + 1938, n = 6; V190G + 1938, n = 6 imaged regions from two independent batches, error bars are s.e.m. k , Representative IF images of day 15 control and mutant (R72W, 1938 1 μM and 1938 2 μM) organoids of BLBP (red). Scale bar, 20 μm. l , Quantification of mean BLBP immunofluorescence intensity of day 15 control and mutant (R72W, 1938 1 μM and 1938 2 μM) organoids. **R72W versus 1938 1 μM, P = 0.0027; **R72W versus 1938 2 μM, P = 0.0037; ****R72W versus control, P = 1.49216 × 10 −6 . Dunnett’s multiple comparison, unpaired two-sided t -test with Welch’s correction. Control + DMSO, n = 6; R72W + DMSO, n = 6; R72W + 1938, n = 5; R72W + 1938, n = 6 imaged regions from two independent batches. Error bars show s.e.m. m , IF images of day 15 control (V190G and DMSO 0.1%) and treated (V190G, UCL-TRO-1938 1 μM) organoids stained for VIM (green) and mitochondrial matrix HSP60 (red). Scale bar, 20 μm. n , Quantification of the nuclear/ cytoplasm ratio of RSL24D1 IF of control + DMSO, AIRIM V190G + DMSO, AIRIM V190G + 1938 2 μM, AIRIM V190G TSC1+/− and control + DMSO, AIRIM R72W + DMSO, AIRIM R72W + 1938 2 μM day 15 cerebral organoids. ****AIRIMV190G versus control, P = 1.26963 × 10 −5 ; * AIRIM V190G versus 1938 2 μM, P = 0.024; ** AIRIM V190G versus AIRIM V190G TSC1 +/−, P = 0.0018; **** AIRIM R72W versus control, P = 1.0138 × 10 −5 ; * AIRIM R72W versus 1938 2 μM, P = 0.0491. Dunnett’s multiple comparison, unpaired two-sided t -test with Welch’s correction. Control (V190G + DMSO), n = 12; V190G + DMSO, n = 2; V190G + 1938 2 μM, n = 6; V190G TSC1+/−, n = 12; control (R72W + DMSO), n = 8; R72W + DMSO, n = 11; V190G + 1938 2 μM, n = 12 organoids from two independent batches. Error bars show s.e.m. o , Model describing a potential explanation for the observed suppression of AIRIM V190G phenotypes by UCL-TRO-1938.

    Techniques Used: Comparison, TUNEL Assay, Control, Mutagenesis, Marker, Immunofluorescence, Staining



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    a , UMAP showing FABP7 expression in control and AIRIM variant cells in day 15 organoids. b , IF image of day 15 control and mutant (V190G and R72W) organoids of transitioning NE marker <t>ZEB2</t> and committed RG marker BLBP (encoded by FABP7). Note that the mutant exhibits less expression of BLBP and less-organized nuclei (marked by ZEB2). Scale bar, 20 µm. c , Quantification of mean BLBP IF intensity of day 15 control and mutant (V190G) organoids. * P = 0.0463. Unpaired two-tailed t -test with Welch’s correction. WT, n = 7; V190G, n = 8 imaged regions from two independent batches. Error bars show s.e.m. d , Quantification of mean BLBP IF intensity of day 15 control and mutant (R72W) organoids. **** P = 8.65486 × 10 −5 . Unpaired two-tailed t -test with Welch’s correction. WT, n = 8; R72W, n = 8 imaged regions from two independent batches. Error bars show s.e.m. e , IF image of day 15 control and AIRIM V190G , AIRIM R72W and AFG2B I466M/V245E organoids of the progenitor marker Sox2 and the neuronal differentiation marker TUBB3. Scale bar, 20 μm. Also included are IF images of day 30 control and AIRIM R72W organoids stained for the progenitor marker Sox2 and the neuronal differentiation marker TUBB3. Scale bar, 40 μm. Three independent batches were performed. f , Representative whole-mount organoid IF images showing the morphology of neural progenitor cells (PAX6 + ), around apical (ZO1 + ) lumens, revealed by sparse labelling with GFP in control and mutant (V190G) organoids. Day 10 cells are columnar and exhibit typical NE morphology. Day 15 control cells show a thinning of apical processes (yellow arrows), whereas mutant cells still seem to be columnar. Scale bar, 20 μm. g , Quantification of the length of neural progenitor cells in control and mutant (V190G) organoids at days 10, 13 and 15, showing reduced length of the progenitor cells in mutant compared with control organoids. Cells with clear apical and basal labelling were used for quantification. Mann–Whitney U -test, **** P < 0.0001, two-tailed, multiple-comparison corrected. Day 10 control, n = 10 cells; day 10 V190G, n = 8 cells; day 13 control, n = 14 cells; day 13 V190G, n = 16 cells; day 15 control, n = 8 cells; and day 15 V190G, n = 8 cells. P adj = 0.0714, day 10; P adj = 9.177792 × 10 −7 , day 13; P adj = 1.098352 × 10 −7 , day 15. Error bars show s.e.m. h , Representative whole-mount organoid IF images showing the morphology of neural progenitor cells revealed by sparse labelling with GFP in control and mutant (R72W) organoids. Day 10 cells are columnar and exhibit typical NE morphology. Day 15 control cells show a thinning of apical processes, whereas mutant cells do not. Scale bar, 20 μm. i , Quantification of the length of neural progenitor cells in control and AIRIM R72W organoids at days 10 and 15. Cells with clear apical and basal labelling were used for quantification. Mann–Whitney U -test, *** P = 0.0009, two-tailed, day 10 control, n = 12 cells; day 10 V190G, n = 9 cells; day 15 control, n = 11 cells; and day 15 V190G, n = 10 cells. Error bars show s.e.m.
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    a , UMAP showing FABP7 expression in control and AIRIM variant cells in day 15 organoids. b , IF image of day 15 control and mutant (V190G and R72W) organoids of transitioning NE marker ZEB2 and committed RG marker BLBP (encoded by FABP7). Note that the mutant exhibits less expression of BLBP and less-organized nuclei (marked by ZEB2). Scale bar, 20 µm. c , Quantification of mean BLBP IF intensity of day 15 control and mutant (V190G) organoids. * P = 0.0463. Unpaired two-tailed t -test with Welch’s correction. WT, n = 7; V190G, n = 8 imaged regions from two independent batches. Error bars show s.e.m. d , Quantification of mean BLBP IF intensity of day 15 control and mutant (R72W) organoids. **** P = 8.65486 × 10 −5 . Unpaired two-tailed t -test with Welch’s correction. WT, n = 8; R72W, n = 8 imaged regions from two independent batches. Error bars show s.e.m. e , IF image of day 15 control and AIRIM V190G , AIRIM R72W and AFG2B I466M/V245E organoids of the progenitor marker Sox2 and the neuronal differentiation marker TUBB3. Scale bar, 20 μm. Also included are IF images of day 30 control and AIRIM R72W organoids stained for the progenitor marker Sox2 and the neuronal differentiation marker TUBB3. Scale bar, 40 μm. Three independent batches were performed. f , Representative whole-mount organoid IF images showing the morphology of neural progenitor cells (PAX6 + ), around apical (ZO1 + ) lumens, revealed by sparse labelling with GFP in control and mutant (V190G) organoids. Day 10 cells are columnar and exhibit typical NE morphology. Day 15 control cells show a thinning of apical processes (yellow arrows), whereas mutant cells still seem to be columnar. Scale bar, 20 μm. g , Quantification of the length of neural progenitor cells in control and mutant (V190G) organoids at days 10, 13 and 15, showing reduced length of the progenitor cells in mutant compared with control organoids. Cells with clear apical and basal labelling were used for quantification. Mann–Whitney U -test, **** P < 0.0001, two-tailed, multiple-comparison corrected. Day 10 control, n = 10 cells; day 10 V190G, n = 8 cells; day 13 control, n = 14 cells; day 13 V190G, n = 16 cells; day 15 control, n = 8 cells; and day 15 V190G, n = 8 cells. P adj = 0.0714, day 10; P adj = 9.177792 × 10 −7 , day 13; P adj = 1.098352 × 10 −7 , day 15. Error bars show s.e.m. h , Representative whole-mount organoid IF images showing the morphology of neural progenitor cells revealed by sparse labelling with GFP in control and mutant (R72W) organoids. Day 10 cells are columnar and exhibit typical NE morphology. Day 15 control cells show a thinning of apical processes, whereas mutant cells do not. Scale bar, 20 μm. i , Quantification of the length of neural progenitor cells in control and AIRIM R72W organoids at days 10 and 15. Cells with clear apical and basal labelling were used for quantification. Mann–Whitney U -test, *** P = 0.0009, two-tailed, day 10 control, n = 12 cells; day 10 V190G, n = 9 cells; day 15 control, n = 11 cells; and day 15 V190G, n = 10 cells. Error bars show s.e.m.

    Journal: Nature Cell Biology

    Article Title: A programmed decline in ribosome levels governs human early neurodevelopment

    doi: 10.1038/s41556-025-01708-8

    Figure Lengend Snippet: a , UMAP showing FABP7 expression in control and AIRIM variant cells in day 15 organoids. b , IF image of day 15 control and mutant (V190G and R72W) organoids of transitioning NE marker ZEB2 and committed RG marker BLBP (encoded by FABP7). Note that the mutant exhibits less expression of BLBP and less-organized nuclei (marked by ZEB2). Scale bar, 20 µm. c , Quantification of mean BLBP IF intensity of day 15 control and mutant (V190G) organoids. * P = 0.0463. Unpaired two-tailed t -test with Welch’s correction. WT, n = 7; V190G, n = 8 imaged regions from two independent batches. Error bars show s.e.m. d , Quantification of mean BLBP IF intensity of day 15 control and mutant (R72W) organoids. **** P = 8.65486 × 10 −5 . Unpaired two-tailed t -test with Welch’s correction. WT, n = 8; R72W, n = 8 imaged regions from two independent batches. Error bars show s.e.m. e , IF image of day 15 control and AIRIM V190G , AIRIM R72W and AFG2B I466M/V245E organoids of the progenitor marker Sox2 and the neuronal differentiation marker TUBB3. Scale bar, 20 μm. Also included are IF images of day 30 control and AIRIM R72W organoids stained for the progenitor marker Sox2 and the neuronal differentiation marker TUBB3. Scale bar, 40 μm. Three independent batches were performed. f , Representative whole-mount organoid IF images showing the morphology of neural progenitor cells (PAX6 + ), around apical (ZO1 + ) lumens, revealed by sparse labelling with GFP in control and mutant (V190G) organoids. Day 10 cells are columnar and exhibit typical NE morphology. Day 15 control cells show a thinning of apical processes (yellow arrows), whereas mutant cells still seem to be columnar. Scale bar, 20 μm. g , Quantification of the length of neural progenitor cells in control and mutant (V190G) organoids at days 10, 13 and 15, showing reduced length of the progenitor cells in mutant compared with control organoids. Cells with clear apical and basal labelling were used for quantification. Mann–Whitney U -test, **** P < 0.0001, two-tailed, multiple-comparison corrected. Day 10 control, n = 10 cells; day 10 V190G, n = 8 cells; day 13 control, n = 14 cells; day 13 V190G, n = 16 cells; day 15 control, n = 8 cells; and day 15 V190G, n = 8 cells. P adj = 0.0714, day 10; P adj = 9.177792 × 10 −7 , day 13; P adj = 1.098352 × 10 −7 , day 15. Error bars show s.e.m. h , Representative whole-mount organoid IF images showing the morphology of neural progenitor cells revealed by sparse labelling with GFP in control and mutant (R72W) organoids. Day 10 cells are columnar and exhibit typical NE morphology. Day 15 control cells show a thinning of apical processes, whereas mutant cells do not. Scale bar, 20 μm. i , Quantification of the length of neural progenitor cells in control and AIRIM R72W organoids at days 10 and 15. Cells with clear apical and basal labelling were used for quantification. Mann–Whitney U -test, *** P = 0.0009, two-tailed, day 10 control, n = 12 cells; day 10 V190G, n = 9 cells; day 15 control, n = 11 cells; and day 15 V190G, n = 10 cells. Error bars show s.e.m.

    Article Snippet: Primary antibodies used in this study include anti-PAX6 (BioLegend, 90130, 1:200 dilution), BLBP (Abcam, ab32423, 1:200 dilution), ZEB2 (Origene, TA802113, 1:150 dilution), ZO1 (BD Biosciences, 610966, 1:300 dilution), GFP (R&D systems, AF4240, 1:200 dilution), HSP60 (ptglab, 15282-1-AP, 1:200 dilution), VIM(V9) (Thermo Fisher, MA5-11883, 1:250 dilution), KI67 (Thermo Fisher, 14-5698-82, 1:100 dilution), p53 (7F5), (Cell Signalling, 2527S, 1:500 dilution), OCT3/4 (SCBT; sc-5279, 1:100 dilution), SOX2(E4) (SCBT, sc-365823, 1:50 dilution) and RSL24D1 (Proteintech, 25190-1-AP, 1:100 dilution).

    Techniques: Expressing, Control, Variant Assay, Mutagenesis, Marker, Two Tailed Test, Staining, MANN-WHITNEY, Comparison

    a , Quantification of bright-field images at day 15. V190G TSC1+/− neural tissue is enlarged relative to V190G TSC1+/+; ** P < 0.01; **** P < 0.0001; Dunnett’s multiple comparison, unpaired two-sided t -test with Welch’s correction. WT TSC1+/+, n = 11; V190G TSC1+/+, n = 12; WT TSC1+/−, n = 37; V190G TSC1+/−, n = 35, from two independent batches; P = 1.734489 × 10 −6 (TSC1+/+ WT versus TSC1+/+ V190G); P = 4.751388 × 10 −6 (TSC1+/+ V190G versus TSC1 +/− V190G); and P = 0.9354378 (TSC1+/− WT versus TSC1+/− V190G). Error bars show s.e.m. b , Representative images showing the TUNEL signal in control TSC1+/+, V190G TSC1+/+, control TSC1+/− and V190G TSC1+/− organoids at day 15. Scale bar, 50 μm. c , Quantification of TUNEL signal of individual NE bud. TUNEL counts were normalized to the area of the imaged bud. **** P < 0.0001, unpaired, two-sided t -test with Welch’s correction. WT TSC1+/+, n = 21; V190G TSC1+/+, n = 22; WT TSC1+/−, n = 17; V190G TSC1+/−, n = 20 NE buds from two independent batches; P = 2.310684 × 10 −7 (TSC1+/+ V190G versus TSC1+/− V190G); and P = 0.1644 (TSC1+/− WT versus TSC1 +/− V190G). Error bars show s.e.m. d , IF image of day 15 control (WT TSC1+/−) and mutant (V190G TSC1+/−) organoids of transitioning NE marker ZEB2 and committed RG marker BLBP. Scale bar, 50 μm. Two independent batch were performed. e , IF image of day 15 control (WT TSC1+/−) and mutant (V190G TSC1+/−) organoids of VIM (green) and mitochondrial matrix HSP60 (red). Scale bar, 20 μm. Two independent batch were performed. f , Quantification of size of day 15 mutant (V190G) organoids treated with vehicle (DMSO) or 1 μM PI3Kα activator UCL-TRO-1938. * P < 0.05. Unpaired two-sided t -test with Welch’s correction. WT + DMSO, n = 7; V190G + DMSO, n = 12; V190G + 1938 1 μM, n = 11, organoids from two independent batches; P = 0.037447. Error bars show s.e.m. g , Quantification of size of day 15 mutant (R72W) organoids treated with vehicle (DMSO) or 1 μM or 2 µM PI3Kα activator UCL-TRO-1938. * P = 0.0373, ** P = 0.0015, **** P = 8.67138 × 10 −7 . Dunnett’s multiple comparison, unpaired t -test with Welch’s correction (two-sided). Control + DMSO, n = 7; R72W + DMSO, n = 6; V190G + 1938 1 μM, n = 6; V190G + 1938 2 μM, n = 6 organoids from two independent batches. Error bars show s.e.m. h , Quantification of TUNEL signal of individual NE bud. TUNEL counts were normalized to the area of the imaged bud. *** P < 0.001, Dunnett’s T3 multiple comparisons test (two-sided). WT + DMSO, n = 9; V190G + DMSO, n = 11; WT + 1938, n = 7; V190G + 1938, n = 13 from two independent batches. P = 0.0008 (V190G + DMSO versus V190G + 1938). Error bars show s.e.m. i , Representative IF images of day 15 control and mutant (V190G, 1938 1 μM and 1938 2 μM) organoids of BLBP (red). Scale bar, 20 μm. j , Quantification of mean BLBP IF intensity of day 15 control and mutant (V190G, 1938 1 μM and 1938 2 μM) organoids. ****V190G versus control, P = 1.60703 × 10 −5 ; ****V190G versus 1938 1 μM, P = 1.39136 × 10 −6 ; ****V190G versus 1938 2 μM, P = 2.35979 × 10 −7 . Dunnett’s multiple comparison unpaired t -test with Welch’s correction (two-sided). Control + DMSO, n = 6; V190G + DMSO, n = 6; control + 1938, n = 6; V190G + 1938, n = 6 imaged regions from two independent batches, error bars are s.e.m. k , Representative IF images of day 15 control and mutant (R72W, 1938 1 μM and 1938 2 μM) organoids of BLBP (red). Scale bar, 20 μm. l , Quantification of mean BLBP immunofluorescence intensity of day 15 control and mutant (R72W, 1938 1 μM and 1938 2 μM) organoids. **R72W versus 1938 1 μM, P = 0.0027; **R72W versus 1938 2 μM, P = 0.0037; ****R72W versus control, P = 1.49216 × 10 −6 . Dunnett’s multiple comparison, unpaired two-sided t -test with Welch’s correction. Control + DMSO, n = 6; R72W + DMSO, n = 6; R72W + 1938, n = 5; R72W + 1938, n = 6 imaged regions from two independent batches. Error bars show s.e.m. m , IF images of day 15 control (V190G and DMSO 0.1%) and treated (V190G, UCL-TRO-1938 1 μM) organoids stained for VIM (green) and mitochondrial matrix HSP60 (red). Scale bar, 20 μm. n , Quantification of the nuclear/ cytoplasm ratio of RSL24D1 IF of control + DMSO, AIRIM V190G + DMSO, AIRIM V190G + 1938 2 μM, AIRIM V190G TSC1+/− and control + DMSO, AIRIM R72W + DMSO, AIRIM R72W + 1938 2 μM day 15 cerebral organoids. ****AIRIMV190G versus control, P = 1.26963 × 10 −5 ; * AIRIM V190G versus 1938 2 μM, P = 0.024; ** AIRIM V190G versus AIRIM V190G TSC1 +/−, P = 0.0018; **** AIRIM R72W versus control, P = 1.0138 × 10 −5 ; * AIRIM R72W versus 1938 2 μM, P = 0.0491. Dunnett’s multiple comparison, unpaired two-sided t -test with Welch’s correction. Control (V190G + DMSO), n = 12; V190G + DMSO, n = 2; V190G + 1938 2 μM, n = 6; V190G TSC1+/−, n = 12; control (R72W + DMSO), n = 8; R72W + DMSO, n = 11; V190G + 1938 2 μM, n = 12 organoids from two independent batches. Error bars show s.e.m. o , Model describing a potential explanation for the observed suppression of AIRIM V190G phenotypes by UCL-TRO-1938.

    Journal: Nature Cell Biology

    Article Title: A programmed decline in ribosome levels governs human early neurodevelopment

    doi: 10.1038/s41556-025-01708-8

    Figure Lengend Snippet: a , Quantification of bright-field images at day 15. V190G TSC1+/− neural tissue is enlarged relative to V190G TSC1+/+; ** P < 0.01; **** P < 0.0001; Dunnett’s multiple comparison, unpaired two-sided t -test with Welch’s correction. WT TSC1+/+, n = 11; V190G TSC1+/+, n = 12; WT TSC1+/−, n = 37; V190G TSC1+/−, n = 35, from two independent batches; P = 1.734489 × 10 −6 (TSC1+/+ WT versus TSC1+/+ V190G); P = 4.751388 × 10 −6 (TSC1+/+ V190G versus TSC1 +/− V190G); and P = 0.9354378 (TSC1+/− WT versus TSC1+/− V190G). Error bars show s.e.m. b , Representative images showing the TUNEL signal in control TSC1+/+, V190G TSC1+/+, control TSC1+/− and V190G TSC1+/− organoids at day 15. Scale bar, 50 μm. c , Quantification of TUNEL signal of individual NE bud. TUNEL counts were normalized to the area of the imaged bud. **** P < 0.0001, unpaired, two-sided t -test with Welch’s correction. WT TSC1+/+, n = 21; V190G TSC1+/+, n = 22; WT TSC1+/−, n = 17; V190G TSC1+/−, n = 20 NE buds from two independent batches; P = 2.310684 × 10 −7 (TSC1+/+ V190G versus TSC1+/− V190G); and P = 0.1644 (TSC1+/− WT versus TSC1 +/− V190G). Error bars show s.e.m. d , IF image of day 15 control (WT TSC1+/−) and mutant (V190G TSC1+/−) organoids of transitioning NE marker ZEB2 and committed RG marker BLBP. Scale bar, 50 μm. Two independent batch were performed. e , IF image of day 15 control (WT TSC1+/−) and mutant (V190G TSC1+/−) organoids of VIM (green) and mitochondrial matrix HSP60 (red). Scale bar, 20 μm. Two independent batch were performed. f , Quantification of size of day 15 mutant (V190G) organoids treated with vehicle (DMSO) or 1 μM PI3Kα activator UCL-TRO-1938. * P < 0.05. Unpaired two-sided t -test with Welch’s correction. WT + DMSO, n = 7; V190G + DMSO, n = 12; V190G + 1938 1 μM, n = 11, organoids from two independent batches; P = 0.037447. Error bars show s.e.m. g , Quantification of size of day 15 mutant (R72W) organoids treated with vehicle (DMSO) or 1 μM or 2 µM PI3Kα activator UCL-TRO-1938. * P = 0.0373, ** P = 0.0015, **** P = 8.67138 × 10 −7 . Dunnett’s multiple comparison, unpaired t -test with Welch’s correction (two-sided). Control + DMSO, n = 7; R72W + DMSO, n = 6; V190G + 1938 1 μM, n = 6; V190G + 1938 2 μM, n = 6 organoids from two independent batches. Error bars show s.e.m. h , Quantification of TUNEL signal of individual NE bud. TUNEL counts were normalized to the area of the imaged bud. *** P < 0.001, Dunnett’s T3 multiple comparisons test (two-sided). WT + DMSO, n = 9; V190G + DMSO, n = 11; WT + 1938, n = 7; V190G + 1938, n = 13 from two independent batches. P = 0.0008 (V190G + DMSO versus V190G + 1938). Error bars show s.e.m. i , Representative IF images of day 15 control and mutant (V190G, 1938 1 μM and 1938 2 μM) organoids of BLBP (red). Scale bar, 20 μm. j , Quantification of mean BLBP IF intensity of day 15 control and mutant (V190G, 1938 1 μM and 1938 2 μM) organoids. ****V190G versus control, P = 1.60703 × 10 −5 ; ****V190G versus 1938 1 μM, P = 1.39136 × 10 −6 ; ****V190G versus 1938 2 μM, P = 2.35979 × 10 −7 . Dunnett’s multiple comparison unpaired t -test with Welch’s correction (two-sided). Control + DMSO, n = 6; V190G + DMSO, n = 6; control + 1938, n = 6; V190G + 1938, n = 6 imaged regions from two independent batches, error bars are s.e.m. k , Representative IF images of day 15 control and mutant (R72W, 1938 1 μM and 1938 2 μM) organoids of BLBP (red). Scale bar, 20 μm. l , Quantification of mean BLBP immunofluorescence intensity of day 15 control and mutant (R72W, 1938 1 μM and 1938 2 μM) organoids. **R72W versus 1938 1 μM, P = 0.0027; **R72W versus 1938 2 μM, P = 0.0037; ****R72W versus control, P = 1.49216 × 10 −6 . Dunnett’s multiple comparison, unpaired two-sided t -test with Welch’s correction. Control + DMSO, n = 6; R72W + DMSO, n = 6; R72W + 1938, n = 5; R72W + 1938, n = 6 imaged regions from two independent batches. Error bars show s.e.m. m , IF images of day 15 control (V190G and DMSO 0.1%) and treated (V190G, UCL-TRO-1938 1 μM) organoids stained for VIM (green) and mitochondrial matrix HSP60 (red). Scale bar, 20 μm. n , Quantification of the nuclear/ cytoplasm ratio of RSL24D1 IF of control + DMSO, AIRIM V190G + DMSO, AIRIM V190G + 1938 2 μM, AIRIM V190G TSC1+/− and control + DMSO, AIRIM R72W + DMSO, AIRIM R72W + 1938 2 μM day 15 cerebral organoids. ****AIRIMV190G versus control, P = 1.26963 × 10 −5 ; * AIRIM V190G versus 1938 2 μM, P = 0.024; ** AIRIM V190G versus AIRIM V190G TSC1 +/−, P = 0.0018; **** AIRIM R72W versus control, P = 1.0138 × 10 −5 ; * AIRIM R72W versus 1938 2 μM, P = 0.0491. Dunnett’s multiple comparison, unpaired two-sided t -test with Welch’s correction. Control (V190G + DMSO), n = 12; V190G + DMSO, n = 2; V190G + 1938 2 μM, n = 6; V190G TSC1+/−, n = 12; control (R72W + DMSO), n = 8; R72W + DMSO, n = 11; V190G + 1938 2 μM, n = 12 organoids from two independent batches. Error bars show s.e.m. o , Model describing a potential explanation for the observed suppression of AIRIM V190G phenotypes by UCL-TRO-1938.

    Article Snippet: Primary antibodies used in this study include anti-PAX6 (BioLegend, 90130, 1:200 dilution), BLBP (Abcam, ab32423, 1:200 dilution), ZEB2 (Origene, TA802113, 1:150 dilution), ZO1 (BD Biosciences, 610966, 1:300 dilution), GFP (R&D systems, AF4240, 1:200 dilution), HSP60 (ptglab, 15282-1-AP, 1:200 dilution), VIM(V9) (Thermo Fisher, MA5-11883, 1:250 dilution), KI67 (Thermo Fisher, 14-5698-82, 1:100 dilution), p53 (7F5), (Cell Signalling, 2527S, 1:500 dilution), OCT3/4 (SCBT; sc-5279, 1:100 dilution), SOX2(E4) (SCBT, sc-365823, 1:50 dilution) and RSL24D1 (Proteintech, 25190-1-AP, 1:100 dilution).

    Techniques: Comparison, TUNEL Assay, Control, Mutagenesis, Marker, Immunofluorescence, Staining

    Journal: Cell

    Article Title: An early cell shape transition drives evolutionary expansion of the human forebrain

    doi: 10.1016/j.cell.2021.02.050

    Figure Lengend Snippet:

    Article Snippet: Primary antibodies used for protein detection, with their corresponding dilutions for immunofluorescence (IF), western blotting (WB) and WB blocking conditions were as follows: mouse anti-β-actin (Abcam, 8226, WB 1:2000 in BSA), mouse anti-ZEB2 (Origene, TA802113, IF 1:150, WB 1:2000 in milk), sheep anti-TBR2 (R&D Systems, AF6166, IF 1:200), mouse anti-CDH2 (BD Biosciences, 610920, IF 1:500, WB 1:1000 in milk), mouse anti-CDH1 (BD Biosciences, 610181, IF 1:500, 1:1000 in milk), rabbit anti-OCLN (Abcam, ab31721, IF 1:200, WB 1:1000 in milk), rabbit anti-EMX1 (ATLAS Antibodies, HPA006421, IF 1:100), rabbit anti-EMX1 (Origene, TA325087, WB 1:1000 in BSA), rabbit anti-BLBP (Abcam, ab32423, IF 1:200), rabbit anti-GLAST (Abcam, ab416, IF 1:200), goat anti-DCX (N-19) (Santa Cruz, sc-8067, IF 1:300), rat anti-CTIP2 (Abcam, ab18465, IF 1:200), mouse anti-HuC/D (Life Technologies, A21271, IF 1:200), mouse anti-ZO1 (BD Biosciences, 610966, IF 1:300), chicken anti-GFP (Thermo Fisher, A10262, IF 1:500), rabbit anti-GFP (Abcam, ab290, WB 1:1000 in milk), rabbit anti-EpCAM (Abcam, ab71916, IF 1:300, WB 1:1000 in milk), mouse anti-Vimentin (V9) (Santa Cruz, sc-6260, IF 1:200, WB 1:1000 in BSA) rabbit anti-PAX6 (Abcam, ab195045, IF 1:200), rabbit anti-SOX2 (Abcam, ab97959, IF 1:200), rabbit anti-SHROOM3 (ATLAS Antibodies, HPA047784, IF 1:200, WB 1:1000), mouse anti-GAPDH (Abcam, ab8245, WB 1:2000 in milk), mouse anti-TUJ1 (Biolegend, 801213, IF 1:500).

    Techniques: Virus, Fluorescence, Recombinant, Knock-Out, Protease Inhibitor, Multiplex Assay, Membrane, Software