vector red chromogen substrate  (Vector Laboratories)

Journal: bioRxiv

Article Title: Tomosyn-2 Regulates Postnatal β-Cell Expansion and Insulin Secretion to Maintain Glucose Homeostasis

doi: 10.1101/2025.05.19.654959

Figure Lengend Snippet: (A) Heatmap showing significantly differentially expressed genes associated with the PI3K–Akt1 signaling pathway in islets of Tomosyn- 2 -/- mice compared to Tomosyn-2 +/+ mice. (B) Representative Western blot showing protein levels of total Akt1, phosphorylated Akt1 (pAkt1), Cyclin D1, and Cdkn1b in islets (n = 3). (C–E) Quantification of Western blot band intensities using ImageJ for (C) total Akt, (D) p-Akt, and (E) Cyclin D1, normalized to β-actin. (F) IF staining of islets from Tomosyn-2 +/+ and Tomosyn-2 -/- mice shows Ki67 (red), insulin (green), and DAPI-stained nuclei (blue). Images captured by confocal microscopy at 20X magnification. Arrows indicate Ki67 insulin + β-cells; insets show magnified views. (G) Quantification of proliferating β-cells (% Ki67 + insulin + β-cells per total insulin+β-cells) using ImageJ (N = 3). (H) Insulin-positive islets were visualized by the presence of blue chromogenic staining in Tomosyn-2 -/- and Tomosyn-2 +/+ mouse pancreata. Quantification of β-cell cross-sectional area (I), circularity (J), β-cell count per islet (K), β-cell number/islet (L), and β-cell size (M). (N) The representative Western blot shows levels of apoptosis marker Caspase-3 in islets (n = 3). (M) Quantification of activated Caspase-3 protein levels, normalized to β-actin. Data are presented as mean ± SEM. An unpaired two-tailed Student’s t-test determined statistical significance. *P < 0.05, **P < 0.01, ***P < 0.001.

Article Snippet: Insulin-positive islets were visualized by the presence of blue chromogenic staining (Vector Labs, Cat #SK-5300).

Techniques: Western Blot, Staining, Confocal Microscopy, Cell Counting, Marker, Two Tailed Test