Journal: bioRxiv

Article Title: A non-canonical AKT1-TERT pathway coordinates autophagy and ERphagy

doi: 10.1101/2025.11.24.690135

Figure Lengend Snippet: Linear regression analysis demonstrating a) a positive correlation between Akt1 and Tert gene expression in Cryba1 cKO mice RPE (n=6) as assessed by qPCR and b) an inverse correlation between Akt2 and Tert gene expression in Cryba1 cKO mice RPE (n=6). c) Western blot analysis demonstrating compensatory AKT isoform signaling in the RPE lysates of Akt2 KI and Akt2 cKO mice (n=3) and d) significant activation of phospho-AKT1 (Ser473) under conditions of AKT2 loss in the RPE of Akt2 cKO mice compared to Akt2 KI. 10μg of protein was loaded per condition, and β-actin was used as loading control. e) qPCR analysis shows significant upregulation of Tert gene expression in the 15-month-old RPE of Akt2 cKO mice, whereas a significant downregulation of Tert expression is observed in the age matched RPE of Akt2 KI mice. f) Telomerase activity measured by Q-TRAP reveals elevated TERT activity in the 15-month-old RPE of Akt2 cKO mice compared to Akt2 KI, while low TERT activity is observed in the age matched RPE of Akt2 KI mice. g) Western blot analysis showing dose-dependent expression of phospho-AKT2 and phospho-AKT1 upon AKT2 inhibition using CCT128930 at varying doses from 6 nM to 100 nM for 48 hours in human fetal RPE cells (hFRPE). The dose range at 6 nM shows the strongest inhibition of AKT2 and compensatory activation of AKT1 in hFRPE cells. 20μg of protein was loaded per condition, and β-actin was used as loading control. h) Line plot showing the differential expression of phospho-AKT1 and phospho-AKT2 upon AKT2 inhibition at varying doses of AKT2 inhibitor (CCT128930). Higher doses above 6 nM show reduced AKT1 compensation and differential expression of phospho-AKT2. Data points represent varying concentrations of the AKT2 inhibitor at log 2 scale of 6 nM, 10 nM, 25 nM, 50 nM, and 100 nM. i) Time course analysis of AKT2 inhibition and AKT1 activation from 30 minutes to 24 hours upon treatment with the AKT2 inhibitor CCT128930 at 6 nM in hFRPE cells. AKT1 activation at the 6-hour time point demonstrates strong TERT phosphorylation at Ser824, which remains stable up to 24 hours. 20μg of protein was loaded per condition, and β-actin was used as loading control. j) Western blot showing changes in the expression of phospho-AKT1 (Ser473), phospho-TERT (Ser824), and compensatory AKT2 activation in telomerase-immortalized RPE cells (hTERT RPE) upon selective AKT1 inhibition using A-674563 HCl at 11 nM, indicating that AKT1 inhibition can significantly impact TERT expression and AKT2 signaling. 20μg of protein was loaded per condition, and Histone H3 (H3) was used as loading control. k) Relative telomerase activity measured using Q-TRAP showed elevated telomerase activity upon AKT2 inhibition at 6 nM in hFRPE cells compared vehicle control while AKT1 inhibition resulted in reduced telomerase activity. RTA analysis was performed using 0.5 µg of lysates per conditions, with HeLa lysates used as a positive control. Values represent mean ± s.d. (n = 3). Statistical analysis was performed using Student’s t -test or one-way ANOVA followed by Tukey’s post hoc test for multiple comparisons. * P < 0.05, ** P < 0.01, *** P < 0.001. ns= no significance

Article Snippet: The primary antibodies Akt1 (2938S), Akt2 (3063S), mTOR (2983S), RPTOR (2280S), RICTOR (2114S), PERK (5683T), p-PERK (3179S), eIF2α (9722S), p-eIF2α (9721S), ATF4 (11815S), BiP (3177T), CHOP (2895T), FOXO3a (12829S), p-FOXO3a (9464S), β-actin (4970S), Vinculin (13901S) and c-MYC (5605T) were purchased from Cell Signaling Technology, Inc. TERT (NB100-317) was purchased from Novus Biologicals.

Techniques: Gene Expression, Western Blot, Activation Assay, Control, Expressing, Activity Assay, Inhibition, Quantitative Proteomics, Phospho-proteomics, Positive Control

Journal: bioRxiv

Article Title: A non-canonical AKT1-TERT pathway coordinates autophagy and ERphagy

doi: 10.1101/2025.11.24.690135

Figure Lengend Snippet: a) Schematic illustrating the experimental design for RNA sequencing analysis of iPSC-derived RPE cells following AKT2 inhibition. Bubble chart plot shows enrichment of b) top ten pathways upregulated pathways, c) top ten downregulated pathways in RNA sequencing analysis of iPSC RPE CFH Y402H treated with AKT2 inhibitor. d) The heatmap shows hierarchical clustering with euclidean distance and Ward’s method of mTOR pathway genes, identifying differentially regulated ( P <0.05, FC>1.5) mTORC1-specific and mTORC2-specific genes in iPSC RPE CFH Y402H cells treated with the AKT2 inhibitor. Upon AKT2 inhibition in iPSC RPE CFH Y402 cells, mTORC2-specific genes are upregulated, while mTORC1-inhibitory genes are DEPTOR and CASTOR2 are upregulated. e) Schematic illustrating the stepwise inhibition strategy utilizing specific inhibitors targeting EGFR, Src, PI3K, mTORC1, mTORC2, and AKT2. f) Western blot analysis displaying the expression of phospho-AKT1, phospho-AKT2, mTOR Ser2448, mTOR Ser2481 and phospho-TERT Ser824 following treatment with inhibitors targeting EGFR (gefitinib; 5μM), Src (PP1; 10μM), PI3K (Wortmannin; 100nM), mTORC1 (Rapamycin; 10nM), mTORC1/2 (Torin1; 50nM) with or without AKT2 inhibition, for 48 hours. A total of 20 µg of protein was loaded in each well, with H3 serving as the loading control. g) Nuclear-cytoplasmic fractionation results demonstrating the expression levels of RPTOR, RICTOR and phosphor-TERT proteins following AKT2 inhibition. A reduction in RPTOR protein levels and an increase in RICTOR protein levels were observed following AKT2 inhibition. A total of 25 µg of protein was loaded in each well. GAPDH was utilized as a control for the cytoplasmic fraction, while Lamin A/C served as a nuclear loading control. h) Correlation analysis indicating an inverse relationship between AKT1 compensation (phospho-AKT1 Ser473) and RPTOR protein levels upon AKT2 inhibition. i) Correlation analysis showing an inverse relationship between AKT1 compensation (phospho-AKT1 Ser473) and RICTOR protein levels upon AKT2 inhibition.

Article Snippet: The primary antibodies Akt1 (2938S), Akt2 (3063S), mTOR (2983S), RPTOR (2280S), RICTOR (2114S), PERK (5683T), p-PERK (3179S), eIF2α (9722S), p-eIF2α (9721S), ATF4 (11815S), BiP (3177T), CHOP (2895T), FOXO3a (12829S), p-FOXO3a (9464S), β-actin (4970S), Vinculin (13901S) and c-MYC (5605T) were purchased from Cell Signaling Technology, Inc. TERT (NB100-317) was purchased from Novus Biologicals.

Techniques: RNA Sequencing, Derivative Assay, Inhibition, Western Blot, Expressing, Control, Fractionation

Journal: bioRxiv

Article Title: A non-canonical AKT1-TERT pathway coordinates autophagy and ERphagy

doi: 10.1101/2025.11.24.690135

Figure Lengend Snippet: a) The schematic displays the C. elegans akt-1 kinase-dead mutant (ΔK222M), like human AKT1 K179M, and trt-1 mutants with AKT1 phosphorylation-deficient mutations (ΔS291A, ΔS355A), comparable to human TERT with AKT1 phosphorylation site mutations (ΔS227A and ΔS824A). qPCR analysis on 15th generation L4 adults (n = 3, 100 worms per condition) shows b) reduced expression of trt-1 (TERT) in akt-1 kinase dead mutant and trt-1 with akt-1 phosphorylation deficient mutants, c) increased expression of akt-1 (AKT1) in akt-1 kinase dead mutant, while reduced expression of akt-1 in trt-1 mutants, d) increased expression of akt-2 in the akt-1 kinase-dead mutant, with a significant elevation noted in the trt-1 mutants. e) Confocal live imaging demonstrates the distribution of functional lysosomes in intestinal epithelial region in the akt-1 kinase-dead and trt-1 mutants, visualized using LysoTracker red and LysoSensor green dye. LysoTracker red highlights active lysosomes, while LysoSensor green dye indicates lysosomal pH changes, providing insights into lysosomal function and integrity. Images reveal altered lysosomal distribution and functionality in mutant strains compared to control groups, emphasizing the impact of AKT-1 kinase activity on lysosomal dynamics. trt-1 variants show low distribution of functional lysosomes compared to akt-1 variants. Scale bar =25μm. f) Quantification of red and yellow puncta reveals significant differences in lysosomal distribution and functionality between mutant strains and wild type groups, highlighting the role of AKT1 kinase activity in regulating lysosomal dynamics. g) Survival line plots show health span of C. elegans measured by pharyngeal pumping for 60 seconds in synchronized L4-stage worms at Days 1, 7 and 15. The trt-1 mutants shows reduced pumping rate and vitality compared to significantly increased pumping rate displayed by akt-1 kinase dead mutant. h) Survival line plot shows percentage lifespan of C elegans variants measured for 30 days. The akt-1 kinase dead mutant shows increased lifespan compared to reduced lifespan in trt-1 mutants. i) Bar graph represents the average lifespan days of each variant. The akt-1 variant has an average lifespan of 23 days comparable to wild type, while trt-1 mutants have a reduced average lifespan of 18 days. j) Schematic illustrates the intestinal region of C. elegans and daf-16 distribution. k) Volumetric analysis showing mean intensity of GFP tagged daf-16 across 57 slices in trt-1 mutant l) Mean intensity of GFP tagged daf-16 across 57 slices in akt-1 mutant. m) Bar graph shows difference in mean intensity of GFP tagged daf-16 in trt-1 and akt-1 mutant. Values represent mean ± s.d. (n = 3). Statistical analysis was performed using Student’s t -test or one-way ANOVA followed by Tukey’s post hoc test for multiple comparisons. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.001. ### P < 0.001. ns= no significance.

Article Snippet: The primary antibodies Akt1 (2938S), Akt2 (3063S), mTOR (2983S), RPTOR (2280S), RICTOR (2114S), PERK (5683T), p-PERK (3179S), eIF2α (9722S), p-eIF2α (9721S), ATF4 (11815S), BiP (3177T), CHOP (2895T), FOXO3a (12829S), p-FOXO3a (9464S), β-actin (4970S), Vinculin (13901S) and c-MYC (5605T) were purchased from Cell Signaling Technology, Inc. TERT (NB100-317) was purchased from Novus Biologicals.

Techniques: Mutagenesis, Phospho-proteomics, Expressing, Imaging, Functional Assay, Control, Activity Assay, Variant Assay

Journal: bioRxiv

Article Title: A non-canonical AKT1-TERT pathway coordinates autophagy and ERphagy

doi: 10.1101/2025.11.24.690135

Figure Lengend Snippet: a) Schematic representation of the experimental design for AKT2 inhibition (CCT128930, 6 nM) in iPSC RPE and hFRPE cells. b) Western blot analysis demonstrates a significant increase in phospho-TERT at Ser824 in the nuclear fraction following AKT2 inhibition in iPSC RPE CFH Y402H cells. The AKT2 inhibition treatment also shows an increase in phospho-AKT1 at Ser473 in the nucleus. This also marked a reduction in nuclear localization of AKT2 upon AKT2 inhibition. A total of 25 µg of protein was loaded in each well. Vinculin was utilized as a control for the cytoplasmic fraction, while Lamin B1 served as a nuclear loading control. c) Densitometry data indicate elevated nuclear phospho-TERT expression following AKT2 inhibition in iPSC RPE CFH Y402H cells. d) Immunofluorescence imaging reveals increased nuclear localization of phospho-TERT after AKT2 inhibition in hFRPE cells. Phalloidin stains the actin cytoskeleton and DAPI staining for the nucleus. (scale bar = 10 µm; zoomed inset = 5 µm). e) Quantification of phospho-TERT intensity in the nuclei of 400 cells (n = 4, with 100 nuclei counted per sample). f) Western blot analysis showing the expression levels of phospho-Src, phospho-Foxo3a, phospho-Foxo1, and PRAS40 in the RPE of 15-month-old Akt2 FL, Akt2 KI, and Akt2 cKO mice (n=3). Elevated Src phosphorylation, indicative of active Src and increased phosphorylation of Foxo3a, which indicates its inactive state in cytoplasm, was observed in the RPE of Akt2 KI mice compared to Akt2 cKO and wild-type controls. A total of 10 µg of protein was loaded in each well, and Vinculin was used as the loading control. g) Western blot analysis shows the subcellular localization of phospho-FOXO3a and total FOXO3a in iPSC RPE CFH Y402H cells. In DMSO-treated controls, phospho-FOXO3a is primarily nuclear, indicating its degradation, while dephosphorylated FOXO3a suggests an active form. Total FOXO3a levels increase in the nucleus after AKT2 inhibition. A total of 25 µg of protein was loaded in each well. β-tubulin was utilized as a control for the cytoplasmic fraction, while H3 served as a nuclear loading control. h) Immunofluorescence imaging reveals increased nuclear localization of FOXO3a after AKT2 inhibition in hFRPE cells. Phalloidin stains the actin cytoskeleton and DAPI staining for the nucleus (scale bar = 10 µm; zoomed inset = 5 µm). i) Quantification of FOXO3a intensity in the nuclei of 400 cells (n = 4, with 100 nuclei counted per sample). Values represent mean ± s.d. (n = 4). Statistical analysis was performed using Student’s t -test or one-way ANOVA followed by Tukey’s post hoc test for multiple comparisons. ** P < 0.01, *** P < 0.001, **** P < 0.001. ns= no significance.

Article Snippet: The primary antibodies Akt1 (2938S), Akt2 (3063S), mTOR (2983S), RPTOR (2280S), RICTOR (2114S), PERK (5683T), p-PERK (3179S), eIF2α (9722S), p-eIF2α (9721S), ATF4 (11815S), BiP (3177T), CHOP (2895T), FOXO3a (12829S), p-FOXO3a (9464S), β-actin (4970S), Vinculin (13901S) and c-MYC (5605T) were purchased from Cell Signaling Technology, Inc. TERT (NB100-317) was purchased from Novus Biologicals.

Techniques: Inhibition, Western Blot, Control, Expressing, Immunofluorescence, Imaging, Staining, Phospho-proteomics

Journal: bioRxiv

Article Title: A non-canonical AKT1-TERT pathway coordinates autophagy and ERphagy

doi: 10.1101/2025.11.24.690135

Figure Lengend Snippet: a) Structural analysis showing the identification of potential allosteric binding pockets for trehalose in the DFG-out conformation of AKT2. Comparative structural models of the activated (PDB: 1O6L) and inactivated (PDB: 8Q61) states demonstrate that only the DFG-out conformation allows binding to the pleckstrin domain, with two candidate binding pockets identified. b) Molecular docking analysis of trehalose binding in pockets 1 and 2, demonstrating favorable binding modes supported by thermodynamically stable water molecules and low energy glycosidic bond torsion values. c) Dose-response analysis of the novel AKT2 inhibitor (nAKT2i) in three independent iPSC-RPE lines, showing sustained suppression of phospho-AKT2 (Ser474) over a broad concentration range (5–100 nM) for up to 48 hours, with a compensatory upregulation of phospho-AKT1 (Ser473). d) Enhanced autophagy flux as indicated by LC3-II and increased clearance of p62 under nAKT2i alone and increased accumulation of p62 when combined with nAKT2i and bafilomycin A in iPSC-RPE CFH Y402H cells demonstrating effective stimulation of autophagic activity. e) Enhanced autophagic process measured by LC3 flux ratio and f) p62 accumulation under nAKT2i with or without bafilomycin A (100nM, 4 hours). Pronounced effects in high-risk AMD polymorphism cells following nAKT2i treatment. g) ERphagy flux was assessed using a tandem GFP-RFP-KDEL construct iPSC-RPE cells with nAKT2i treatment, emphasizing the inhibitor’s role in targeting damaged ER components for degradation. h) Quantification shows an increase in the RFP puncta ratio upon AKT2 inhibition indicate a shift towards autolysosome formation and enhanced ERphagy flux. i) Activation of both the PERK and IRE1α branches of the UPR mechanism in a ‘disease in a dish’ model after nAKT2i treatment, leading to reduced ER stress, as shown by decreased levels of CHOP compared to CC-HS alone conditions. Densitometry analysis showing normalized expression of j) phospho-PERK, k) IRE1α, l) BiP, m) PDI and n) CHOP. o) Western blot showing expression of ApoE in following nAKT2i treatment in the presence of CC-HS, p) comparative expression levels of ApoE in iPSC-RPE cells, highlighting the reduction of ApoE levels with nAKT2i q) Enhanced expression of the ER-phagy receptor CCPG1 in iPSC RPE cells treated with nAKT2i compared to CC-HS alone, r) comparative expression levels of CCPG1 in iPSC-RPE cells, suggesting its potential role in alleviating ER stress and promoting cellular homeostasis. Values represent mean ± s.d. (n = 4). Statistical analysis was performed using one-way ANOVA followed by Tukey’s post hoc test for multiple comparisons. * P < 0.05, ** P < 0.01, **** P < 0.0001.

Article Snippet: The primary antibodies Akt1 (2938S), Akt2 (3063S), mTOR (2983S), RPTOR (2280S), RICTOR (2114S), PERK (5683T), p-PERK (3179S), eIF2α (9722S), p-eIF2α (9721S), ATF4 (11815S), BiP (3177T), CHOP (2895T), FOXO3a (12829S), p-FOXO3a (9464S), β-actin (4970S), Vinculin (13901S) and c-MYC (5605T) were purchased from Cell Signaling Technology, Inc. TERT (NB100-317) was purchased from Novus Biologicals.

Techniques: Binding Assay, Concentration Assay, Activity Assay, Construct, Inhibition, Activation Assay, Expressing, Western Blot

Journal: Oncology reports

Article Title: Anti‑oncogenic and pro‑myogenic action of the MKK6/p38/AKT axis induced by targeting MEK/ERK in embryonal rhabdomyosarcoma.

doi: 10.3892/or.2022.8363

Figure Lengend Snippet: Figure 8. AKT is induced by MEK/ERK inhibition. RD and TE cells were treated with 10 µM U0126 or left untreated and lysates were analysed for AKT‑PO4 Ser473 and AKT‑PO4 Thr308 expression. Both phosphorylation levels were increased after O/N, 1 day and 3 days of treatments. The numbers on the left of the blots indicate the protein size (kDa). In the lower panels, quantitative evaluations of the western blots are shown as the mean ± SD. Statistical analyses were performed using a Student's t‑test: ***P<0.001; **P<0.01; *P<0.05 vs. negative control (control). Experiments were performed twice. TE cells, TE671 cells; O/N, overnight.

Article Snippet: Filters were blocked with 5% non‐fat dry milk or 3% BSA for 1 h at room temperature and incubated overnight at 4 ̊C with the following primary anti‐ bodies: Anti‐Myc (cat. no. sc‐40; 1:300), anti‐ERK‐PO4 E‐4 (cat. no. sc‐7383; 1:500), anti‐ERK1/2 C‐9 (cat. no. sc‐514302; 1:500), anti‐p38‐PO4 (cat. no. sc‐166182; 1:1,000), anti‐p38 (cat. no. sc‐535; 1:500), anti‐MKK3 (cat. no. sc‐961; 1:500), anti‐cyclin D1 (cat. no. sc‐20044; 1:1,000), anti‐p21 (cat. no. sc‐6246; 1:200), anti‐GAPDH (cat. no. sc‐47724; 1:500) and anti‐tubulin (cat. no. sc‐5286; 1:500) (all from Santa Cruz Biotechnology, Inc.); anti MKK6‐PO4 D8E9 (MEK3/6) (cat. no. 12280; 1:1,000), anti‐MKK6 D31D1 (cat. no. 8550; 1:1,000), anti‐AKT‐PO4 Thr308 (cat. no. 4056; 1:1,000), anti‐AKT‐PO4 Ser473 (cat. no. 9271; 1:1,000) and AKT (cat. no. 9272; 1:1,000) (all from Cell Signalling Technology, Inc.); anti‐myosin heavy chain (MHC; cat. no. MF20; 1:300) and anti‐myogenin (cat. no. F5D; 1:300) were monoclonal from hybridoma supernatant (all from Developmental Studies Hybridoma Bank).

Techniques: Inhibition, Expressing, Phospho-proteomics, Western Blot, Negative Control, Control

Journal: Oncology reports

Article Title: Anti‑oncogenic and pro‑myogenic action of the MKK6/p38/AKT axis induced by targeting MEK/ERK in embryonal rhabdomyosarcoma.

doi: 10.3892/or.2022.8363

Figure Lengend Snippet: Figure 9. AKT activation is part of myogenic differentiation in RD cells. (A) Cells transfected with empty vector (CMV), caMKK3 or caMKK6 were analysed for AKT‑PO4 Ser473, AKT‑PO4 Thr308 and p38‑PO4 expression. Both AKT phosphorylation sites and p38 were activated by caMKK6 transfection, whereas they were absent in CMV and caMKK3 transfected cells; MKK3 and MKK6 overexpression is shown as a transfection control. GAPDH was used for protein quantification. Phospho‑kinases were also normalised for unphosphorylated isoforms. The numbers on the left of the blots indicate the protein size (kDa). Right panel represents histograms of the quantitative evaluations of the western blots, expressed as the mean ± SD. Statistical analyses were performed using one‑way ANOVA with Dunnett's post hoc test: ***P<0.001 vs. CMV. (B) Western blots showing the reduced AKT phosphorylation in caMKK6‑transfected RD cells treated with 5 µM SB203580. (C) Both AKT phosphorylation levels were not activated by U0126 in p38‑silenced RD cells, whilst they were present in scramble control‑transfected (SCR) cells treated with U0126. GAPDH was used as loading control. (B and C) The numbers on the left of the blots indicate the protein size (kDa). Lower panels represent histograms of the quantitative evaluations of the western blots, expressed as the mean ± SD. Statistical analyses were performed using two‑way ANOVA with Tukey's post hoc test: ***P<0.001; **P<0.01; *P<0.05 vs. CMV or SCR; ###P<0.001 vs. caMKK6; §§§P<0.001 vs. SCR‑U0126. Experiments were performed twice.

Article Snippet: Filters were blocked with 5% non‐fat dry milk or 3% BSA for 1 h at room temperature and incubated overnight at 4 ̊C with the following primary anti‐ bodies: Anti‐Myc (cat. no. sc‐40; 1:300), anti‐ERK‐PO4 E‐4 (cat. no. sc‐7383; 1:500), anti‐ERK1/2 C‐9 (cat. no. sc‐514302; 1:500), anti‐p38‐PO4 (cat. no. sc‐166182; 1:1,000), anti‐p38 (cat. no. sc‐535; 1:500), anti‐MKK3 (cat. no. sc‐961; 1:500), anti‐cyclin D1 (cat. no. sc‐20044; 1:1,000), anti‐p21 (cat. no. sc‐6246; 1:200), anti‐GAPDH (cat. no. sc‐47724; 1:500) and anti‐tubulin (cat. no. sc‐5286; 1:500) (all from Santa Cruz Biotechnology, Inc.); anti MKK6‐PO4 D8E9 (MEK3/6) (cat. no. 12280; 1:1,000), anti‐MKK6 D31D1 (cat. no. 8550; 1:1,000), anti‐AKT‐PO4 Thr308 (cat. no. 4056; 1:1,000), anti‐AKT‐PO4 Ser473 (cat. no. 9271; 1:1,000) and AKT (cat. no. 9272; 1:1,000) (all from Cell Signalling Technology, Inc.); anti‐myosin heavy chain (MHC; cat. no. MF20; 1:300) and anti‐myogenin (cat. no. F5D; 1:300) were monoclonal from hybridoma supernatant (all from Developmental Studies Hybridoma Bank).

Techniques: Activation Assay, Transfection, Plasmid Preparation, Expressing, Phospho-proteomics, Over Expression, Control, Western Blot

Journal: The Journal of endocrinology

Article Title: Growth factor-induced signaling of the pancreatic epithelium.

doi: 10.1677/joe.1.05949

Figure Lengend Snippet: Figure 5 Phospho-Akt and Akt staining in the IFN transgenic pancreas. Pancreatic sections from NOD (A and C) and IFNNOD (B and D). (A and B) Sections stained with an antibody to phospho-Akt (Ser473). A significant number of cells in the proliferating ducts and cells contiguous to the duct cells exhibit phospho-Akt expression in the transgenic mouse pancreas. Interestingly, phospho-Akt is also expressed in the periphery of the islets in the NOD pancreas in a pattern similar to glucagon staining. (C and D) Sections stained with an antibody to Akt. Original magnification 50.

Article Snippet: The polyclonal phospho-Akt antibodies were generated against a synthetic phospho-Ser473 peptide corresponding to residues around mouse Akt (Cell Signaling Technology).

Techniques: Staining, Transgenic Assay, Expressing