mab1556  (R&D Systems)

Journal: Nature Physics

Article Title: Boundary-guided cell alignment drives mouse epiblast maturation

doi: 10.1038/s41567-026-03176-9

Figure Lengend Snippet: ( A ) Box plots of EPI cell number, showing median (centre), 25th–75th percentiles (box), with whiskers extending to 1.5x interquartile range, with individual embryos shown as dots. Sample sizes: Lamc1 +/+ , n = 9 (E4.5), 13 (E4.75), 11 (E5.0); Lamc1 +/− , n = 12, 28, 20; Lamc1 −/− , n = 4, 11, 9 embryos from 5, 7, and 5 independent Lamc1 +/− × Lamc1 +/− litters. One-way ANOVA with Tukey’s post hoc test, ns = not significant. ( B ) Box plots of EPI cell number, as in ( A ). Sample sizes: Itgb1 +/+ , n = 5 (E4.5), 13 (E4.75), 9 (E5.0); Itgb1 +/− , n = 13, 28, 26; Itgb1 −/− , n = 8, 14, 11 embryos from 5, 8, and 7 independent Itgb1 +/− × Itgb1 +/− litters. One-way ANOVA with Tukey’s post hoc test. *p < 0.05, **p < 0.01, ns = not significant. At E5.0, Itgb1 +/+ vs Itgb1 −/− , p = 0.017; Itgb1 +/− vs Itgb1 −/− , p = 0.001. ( C ) Immunofluorescence images of representative Lamc1 +/+ or +/− and Lamc1 −/− embryos at E5.25, stained for Oct3/4 (EPI, cyan), phalloidin (yellow), and podocalyxin (magenta). n = 28 ( Lamc1 +/+ or +/− ), 6 ( Lamc1 −/− ) embryos from 4 independent Lamc1 +/− × Lamc1 +/− litters. ( D ) Immunofluorescence images of representative Itgb1 +/+ or +/− and Itgb1 −/− embryos at E5.25, stained for Oct3/4 (EPI, cyan), pan-laminin (green), and phalloidin (yellow). n = 13 ( Itgb1 +/+ or +/− ), 2 ( Itgb1 −/− ) embryos from 2 independent Itgb1 +/− × Itgb1 +/− litters. ( E ) Long axis length measurement, shown as violin plots with individual data points. Dot colours indicate embryo stage; red bars show medians. Sample sizes: Lamc1 +/+ , n = 163 cells from 6 embryos (E4.5), 397 from 8 (E4.75), 571 from 10 (E5.0); Lamc1 +/− , n = 199 from 7, 583 from 14, 637 from 10; Lamc1 − / − , n = 75 from 3, 277 from 7, 462 from 9. Mann-Whitney U test (two-sided) without correction for multiple comparisons, each group compared to the reference group [15-29 cells]. *p < 0.05, **p < 0.01, ***p < 0.001, ns = not significant. Exact p-values: Lamc1 +/+ [30–44], p = 0.345; [45–59], p = 0.189; [60–74], p = 0.236; [75–89], p = 8.72 × 10 −9 . Lamc1 +/− [30–44], p = 0.051; [45–59], p = 1.00 × 10 −3 ; [60–74], p = 5.02 × 10 −6 ; [75–89], p = 7.42 × 10 −6 . Lamc1 −/− [30–44], p = 0.539; [45–59], p = 4.07 × 10 −4 ; [60–74], p = 0.059. ( F ) Long axis length measurement, as in ( E ). Sample sizes: Itgb1 +/+ , n = 110 cells from 3 embryos (E4.5), 399 from 8 (E4.75), 251 from 4 (E5.0); Itgb1 +/− , n = 200 from 5, 560 from 11, 618 from 10; Itgb1 −/− , n = 209 from 7, 470 from 11, 360 from 9. Mann-Whitney U test (two-sided) without correction for multiple comparisons, each group compared to the reference group [15-29 cells]. *p < 0.05, **p < 0.01, ***p < 0.001, ns = not significant. Exact p-values: Itgb1 +/+ : [30–44], p = 2.36 × 10 −4 ; [45–59], p = 0.015; [60–74], p = 8.97 × 10 −3 ; [75–89], p = 2.34 × 10 −4 . Itgb1 +/− : [30–44], p = 0.845; [45–59], p = 0.335; [60–74], p = 0.518; [75–89], p = 0.022. Itgb1 −/− : [30–44], p = 5.65 × 10 −4 ; [45–59], p = 0.022; [60–74], p = 3.71 × 10 −4 . Scale bars, 20 µm. See also Fig. .

Article Snippet: Primary antibodies against laminin (Novus-Biologicals, NB300-144), pERM (Cell Signaling, 3726) and podocalyxin (R&D Systems, MAB1556) were diluted at 1:200.

Techniques: Immunofluorescence, Staining, MANN-WHITNEY

Journal: Advanced Science

Article Title: Calcium Shock Enables Efficient and Programmable Particle Delivery for Genome Editing Applications

doi: 10.1002/advs.202510441

Figure Lengend Snippet: CaSh‐Pro enables programmable delivery into specific cell types in human organoids. (A) General schematic of CaSh‐Pro approach in heterocellular epithelia. (B) Detailed illustration of CaSh‐Pro mechanism in the kidney organoid structure. Cas9 complexes tethered to antibodies bind to specific cell types and are endocytosed, enabling genome editing in organoids. (C) Molecular illustration of Cas9 enzyme complex used as the basis for CaSh‐Pro. Cas9 RNP in blue/white, fused protein G domain in orange, and IgG MTA in red. The model is based on crystal structures determined for the constituent components, with respective Protein Data Bank IDs 4ZT0, 1FCC, and 1HZH. (D) Confocal live and (E) fixed fluorescence images of PodoTracker (αPODXL) or TubuleTracker (αECAD), with podocyte (WT1) and tubule (LTL) surrogate markers. (F) Quantification of the percentage of PodoTracker or TubuleTracker overlapping with PODXL + , LTL + , or other cell types (mean ± s.e.m., n=9 organoids pooled from n = 3 independent biological experiments; ns : not significant, **** , p < 0.0001, Ordinary one‐way ANOVA with multiple comparisons). (G) Confocal immunofluorescence images of kidney organoids under different genome editing treatment conditions, compared to untreated. Transfection of CRISPR RNP was conducted with the endosomolytic peptide E5TAT ± PodoTracker antibody. (H) Quantification of % TdTomato + cells expressing the podocyte marker WT1 (mean ± s.e.m., n = 17 organoids per condition, pooled from three independent biological experiments, ns : not significant, **** , p < 0.0001, Ordinary one‐way ANOVA with multiple comparisons). (I,J) As for (G,H), but using TubuleTracker instead of PodoTracker, and the proximal tubule marker LTL instead of WT1 (mean ± s.e.m., n = 14–19 organoids per condition, pooled from three independent biological experiments, ns : not significant, **** , p < 0.0001, Ordinary one‐way ANOVA with multiple comparisons).

Article Snippet: Primary antibodies and labels included PODXL (R&D system AF1658, 1:400), biotinylated LTL (Vector Labs B‐1325, 1:400), WT1 (Abcam ab89901, 1:500), ECAD (Abcam ab11512, 1:500), gamma H2AX (Cell Signaling 9718S, 1:500), APOL1 (Genentech A.17A5, 1:300), β‐Tubulin III (Sigma Aldrich, T2200), MLC2A (Synaptic System, 311011), CD144 (BD Biosciences 555661, 1:300), MUC1 (Abcam ab15481, 1:200), P63 (R&D BAF1916, 1:40) and NKX2.1 (Seven Hills Bioreagents WRAB‐1231, 1:500).

Techniques: Fluorescence, Immunofluorescence, Transfection, CRISPR, Expressing, Marker