Journal: Scientific Reports

Article Title: Elevated ITGA3 expression serves as a novel prognostic biomarker and regulates tumor progression in cervical cancer

doi: 10.1038/s41598-024-75770-x

Figure Lengend Snippet: Aberrant expression of ITGA3 in different tumors. ( A ) The levels of ITGA3 in human cancers. ( B ) Univariate cox analysis of ITGA3 expression in different tumors from TCGA and overall survival. ( C - F ) Relationship between ITGA3 expression and overall survival ( C ), disease-free survival ( D ), disease-free interval ( E ) and progression-free interval ( F ) in cervical cancer. ( G ) The HE staining image of normal cervical tissue and cervical cancer. ( H ) The validation of ITGA3 expression in GEO datasets. ( I ) Using serial sections to do immunohistochemical experiments on both HE and ITGA3. *** p < 0.001, ** p < 0.01, * p < 0.05.

Article Snippet: Then, the membranes were incubated with primary antibodies against ITGA3 (1:1000; ABclonal, China, Cat: A17502), phospho-AKT-Ser473 (1:1,000; Affinity, China, Cat: #AF0016), phospho-PI3K-p85 (1:1000; Affinity, China, Cat: #AF3242), BAX (1:1000; ABclonal, China, Cat: A0207), Bcl2 (1:1000; ABclonal, China, Cat: A19693), E-cadherin (1:50000; proteintech, China, Cat: A20874-1-AP), N-cadherin (1:1000; ABclonal, China, Cat: A19083), vimentin (1:1000; ABclonal, China, Cat: A19607), MMP2(1:1000; ABclonal, China, Cat: A11144), MMP9 (1:1000; ABclonal, China, Cat: A11147) or GAPDH (1:10000, ABclonal, China, Cat: AC036) overnight at 4 °C.

Techniques: Expressing, Staining, Biomarker Discovery, Immunohistochemical staining

Journal: Scientific Reports

Article Title: Elevated ITGA3 expression serves as a novel prognostic biomarker and regulates tumor progression in cervical cancer

doi: 10.1038/s41598-024-75770-x

Figure Lengend Snippet: Immune cell infiltration in ITGA3 groups. ( A ) The relationship between immune cells and the levels of ITGA3 using the XCELL, TIMER, QUANTISEQ, MCPCOUNTER, EPIC, CIBERSORT-ABS and CIBERSORT algorithm. ( B ) The correlation analysis between ITGA3 expression and immune cell infiltration. ( C ) The abundance of tumor immune infiltrating cell between two risk groups. *** p < 0.001, ** p < 0.01, * p < 0.05. ns means no statistical difference.

Article Snippet: Then, the membranes were incubated with primary antibodies against ITGA3 (1:1000; ABclonal, China, Cat: A17502), phospho-AKT-Ser473 (1:1,000; Affinity, China, Cat: #AF0016), phospho-PI3K-p85 (1:1000; Affinity, China, Cat: #AF3242), BAX (1:1000; ABclonal, China, Cat: A0207), Bcl2 (1:1000; ABclonal, China, Cat: A19693), E-cadherin (1:50000; proteintech, China, Cat: A20874-1-AP), N-cadherin (1:1000; ABclonal, China, Cat: A19083), vimentin (1:1000; ABclonal, China, Cat: A19607), MMP2(1:1000; ABclonal, China, Cat: A11144), MMP9 (1:1000; ABclonal, China, Cat: A11147) or GAPDH (1:10000, ABclonal, China, Cat: AC036) overnight at 4 °C.

Techniques: Expressing

Journal: Scientific Reports

Article Title: Elevated ITGA3 expression serves as a novel prognostic biomarker and regulates tumor progression in cervical cancer

doi: 10.1038/s41598-024-75770-x

Figure Lengend Snippet: The expression of immune checkpoints in the ITGA3 groups. ( A ) The expression of immune checkpoints among two risk groups and normal cervical tissue. ( B - H ) The abundance of tumor immune infiltrating cell between two risk groups. ( I - L ) The score of CD8 + T cell infiltration ( I ), CAF ( J ), MDSC ( K ), TAM ( L ) infiltration in two risk groups using TIDE algorithm. *** p < 0.001, ** p < 0.01, * p < 0.05. ns means no statistical difference. CAF: cancer associated fibroblasts; MDSC: marrow derived suppressor cells; TAM: tumor associated macrophage; TIDE: tumor immune dysfunction and exclusion.

Article Snippet: Then, the membranes were incubated with primary antibodies against ITGA3 (1:1000; ABclonal, China, Cat: A17502), phospho-AKT-Ser473 (1:1,000; Affinity, China, Cat: #AF0016), phospho-PI3K-p85 (1:1000; Affinity, China, Cat: #AF3242), BAX (1:1000; ABclonal, China, Cat: A0207), Bcl2 (1:1000; ABclonal, China, Cat: A19693), E-cadherin (1:50000; proteintech, China, Cat: A20874-1-AP), N-cadherin (1:1000; ABclonal, China, Cat: A19083), vimentin (1:1000; ABclonal, China, Cat: A19607), MMP2(1:1000; ABclonal, China, Cat: A11144), MMP9 (1:1000; ABclonal, China, Cat: A11147) or GAPDH (1:10000, ABclonal, China, Cat: AC036) overnight at 4 °C.

Techniques: Expressing, Derivative Assay

Journal: Scientific Reports

Article Title: Elevated ITGA3 expression serves as a novel prognostic biomarker and regulates tumor progression in cervical cancer

doi: 10.1038/s41598-024-75770-x

Figure Lengend Snippet: Co-expression network of ITGA3 and correlated pathway analysis. ( A ) The network of binding proteins of ITGA3 by STRING website. ( B ) The impact of ITGA3-related genes on tumor signaling pathways in pan-cancer by GSCALite websites. ( C ) The impact of ITGA3-related genes on tumor signaling pathways in cervical cancer. ( D ) The enrichment score of tumor progression and tumor metabolism between two risk groups in cervical cancer. *** p < 0.001, ** p < 0.01. ns means no statistical difference.

Article Snippet: Then, the membranes were incubated with primary antibodies against ITGA3 (1:1000; ABclonal, China, Cat: A17502), phospho-AKT-Ser473 (1:1,000; Affinity, China, Cat: #AF0016), phospho-PI3K-p85 (1:1000; Affinity, China, Cat: #AF3242), BAX (1:1000; ABclonal, China, Cat: A0207), Bcl2 (1:1000; ABclonal, China, Cat: A19693), E-cadherin (1:50000; proteintech, China, Cat: A20874-1-AP), N-cadherin (1:1000; ABclonal, China, Cat: A19083), vimentin (1:1000; ABclonal, China, Cat: A19607), MMP2(1:1000; ABclonal, China, Cat: A11144), MMP9 (1:1000; ABclonal, China, Cat: A11147) or GAPDH (1:10000, ABclonal, China, Cat: AC036) overnight at 4 °C.

Techniques: Expressing, Binding Assay, Protein-Protein interactions

Journal: Scientific Reports

Article Title: Elevated ITGA3 expression serves as a novel prognostic biomarker and regulates tumor progression in cervical cancer

doi: 10.1038/s41598-024-75770-x

Figure Lengend Snippet: Functional enrichment pathway analysis. ( A and B ) GO and KEGG pathways based on ITGA3 binding proteins and interactive genes. ( C ) Recognizing different gene sets in the high- and low-risk groups with GSEA analysis.

Article Snippet: Then, the membranes were incubated with primary antibodies against ITGA3 (1:1000; ABclonal, China, Cat: A17502), phospho-AKT-Ser473 (1:1,000; Affinity, China, Cat: #AF0016), phospho-PI3K-p85 (1:1000; Affinity, China, Cat: #AF3242), BAX (1:1000; ABclonal, China, Cat: A0207), Bcl2 (1:1000; ABclonal, China, Cat: A19693), E-cadherin (1:50000; proteintech, China, Cat: A20874-1-AP), N-cadherin (1:1000; ABclonal, China, Cat: A19083), vimentin (1:1000; ABclonal, China, Cat: A19607), MMP2(1:1000; ABclonal, China, Cat: A11144), MMP9 (1:1000; ABclonal, China, Cat: A11147) or GAPDH (1:10000, ABclonal, China, Cat: AC036) overnight at 4 °C.

Techniques: Functional Assay, Binding Assay

Journal: Scientific Reports

Article Title: Elevated ITGA3 expression serves as a novel prognostic biomarker and regulates tumor progression in cervical cancer

doi: 10.1038/s41598-024-75770-x

Figure Lengend Snippet: The impact of ITGA3 on t apoptosis rates in cervical cancer cell. ( A ) The immunofluorescence images of ITGA3 in human cells from the HPA database. Blue: cell nucleus, red: microtubules, and green: ITGA3. ( B ) The expression of ITGA3 in various cervical cancer cell lines in CCLE database. ( C and D ) WB analysis of ITGA3 in cells transfected with ov-NC, si-NC, ov-ITGA3 and si-ITGA3 lentivirus. ( E ) PCR analysis of ITGA3 in ov-NC, si-NC, ov-ITGA3 and si-ITGA3 groups. ( F ) Cell viability in ov-NC, si-NC, ov-ITGA3 and si-ITGA3 groups detected by CCK-8 assay. p represents the comparison with the si-NC group, while # represents the comparison with the ov-NC group. ** p < 0.01, * p < 0.05. ## p < 0.01, # p < 0.05 ns means no statistical difference. (G and H) Flow cytometry assay displayed the apoptosis rates in four groups and bar charts are used for visualization. (K-M) WB analysis of Bax and Bcl-2 protein expression in two groups. *** p < 0.001, ** p < 0.01, * p < 0.05. ns means no statistical difference.

Article Snippet: Then, the membranes were incubated with primary antibodies against ITGA3 (1:1000; ABclonal, China, Cat: A17502), phospho-AKT-Ser473 (1:1,000; Affinity, China, Cat: #AF0016), phospho-PI3K-p85 (1:1000; Affinity, China, Cat: #AF3242), BAX (1:1000; ABclonal, China, Cat: A0207), Bcl2 (1:1000; ABclonal, China, Cat: A19693), E-cadherin (1:50000; proteintech, China, Cat: A20874-1-AP), N-cadherin (1:1000; ABclonal, China, Cat: A19083), vimentin (1:1000; ABclonal, China, Cat: A19607), MMP2(1:1000; ABclonal, China, Cat: A11144), MMP9 (1:1000; ABclonal, China, Cat: A11147) or GAPDH (1:10000, ABclonal, China, Cat: AC036) overnight at 4 °C.

Techniques: Immunofluorescence, Expressing, Transfection, CCK-8 Assay, Comparison, Flow Cytometry

Journal: Scientific Reports

Article Title: Elevated ITGA3 expression serves as a novel prognostic biomarker and regulates tumor progression in cervical cancer

doi: 10.1038/s41598-024-75770-x

Figure Lengend Snippet: Increment of ITGA3 promotes tumor progression in vitro. ( A - D ) wound healing ( A and B ) and transwell assays ( C and D ) were used to detect the ability of cell migration and invasion between ov-NC and ov-ITGA3 groups. ( E - K ) WB analysis of N-cadherin, E-cadherin, vimentin, MMP2 and MMP9 in two groups. ( L - N ) WB analysis of p-I3K and p-Akt in two groups. Data are shown as means ± S.D. * p < 0.05, ** p < 0.01, *** p < 0.001. ns means no statistical difference.

Article Snippet: Then, the membranes were incubated with primary antibodies against ITGA3 (1:1000; ABclonal, China, Cat: A17502), phospho-AKT-Ser473 (1:1,000; Affinity, China, Cat: #AF0016), phospho-PI3K-p85 (1:1000; Affinity, China, Cat: #AF3242), BAX (1:1000; ABclonal, China, Cat: A0207), Bcl2 (1:1000; ABclonal, China, Cat: A19693), E-cadherin (1:50000; proteintech, China, Cat: A20874-1-AP), N-cadherin (1:1000; ABclonal, China, Cat: A19083), vimentin (1:1000; ABclonal, China, Cat: A19607), MMP2(1:1000; ABclonal, China, Cat: A11144), MMP9 (1:1000; ABclonal, China, Cat: A11147) or GAPDH (1:10000, ABclonal, China, Cat: AC036) overnight at 4 °C.

Techniques: In Vitro, Migration

Journal: Scientific Reports

Article Title: Elevated ITGA3 expression serves as a novel prognostic biomarker and regulates tumor progression in cervical cancer

doi: 10.1038/s41598-024-75770-x

Figure Lengend Snippet: Loss of ITGA3 attenuates tumor progression in vitro. ( A - D ) wound healing (A and B) and transwell assays ( C and D ) were used to detect the ability of cell migration and invasion between si-NC and si-ITGA3 groups. ( E - K ) WB analysis of N-cadherin, E-cadherin, vimentin, MMP2 and MMP9 in two groups. ( L - N ) WB analysis of p-I3K and p-Akt in two groups. Data are shown as means ± S.D. * p < 0.05, ** p < 0.01, *** p < 0.001. ns means no statistical difference.

Article Snippet: Then, the membranes were incubated with primary antibodies against ITGA3 (1:1000; ABclonal, China, Cat: A17502), phospho-AKT-Ser473 (1:1,000; Affinity, China, Cat: #AF0016), phospho-PI3K-p85 (1:1000; Affinity, China, Cat: #AF3242), BAX (1:1000; ABclonal, China, Cat: A0207), Bcl2 (1:1000; ABclonal, China, Cat: A19693), E-cadherin (1:50000; proteintech, China, Cat: A20874-1-AP), N-cadherin (1:1000; ABclonal, China, Cat: A19083), vimentin (1:1000; ABclonal, China, Cat: A19607), MMP2(1:1000; ABclonal, China, Cat: A11144), MMP9 (1:1000; ABclonal, China, Cat: A11147) or GAPDH (1:10000, ABclonal, China, Cat: AC036) overnight at 4 °C.

Techniques: In Vitro, Migration

Journal: Life Science Alliance

Article Title: Hypoxia favors tumor growth in colorectal cancer in an integrin αDβ1/hemoglobin δ-dependent manner

doi: 10.26508/lsa.202402925

Figure Lengend Snippet: Integrin and erythroid cell genes expressed in Colo205 cells grown in hypoxia.

Article Snippet: The primers for Hbβ, Hbδ, and αD integrin were obtained from Origene.

Techniques:

Journal: Life Science Alliance

Article Title: Hypoxia favors tumor growth in colorectal cancer in an integrin αDβ1/hemoglobin δ-dependent manner

doi: 10.26508/lsa.202402925

Figure Lengend Snippet: (A) Cells were stained for integrin αD (green, rabbit polyclonal antibody), and β2 integrin (red) chains showed absence of β2. Integrin β1 staining (red) was strong and it co-distributed with αD staining. Scale bar 10 μm. (B) Cells were grown in normoxia, transferred to hypoxia, and stained at different time points for αD (red) and Hb (green, goat polyclonal). After 2 h under hypoxia, the cells showed increased staining for αD, but weak staining for Hb. After 9 h, Hb staining was strong and showed partial co-distribution with αD. After 24 h, cells expressed cup-like structures. Scale bar 10 μm. The last figure shows the integrin αD (red) and Hbδ (green) at 24 h under hypoxic conditions. The figure was obtained from the . Scale bar 10 μm. (C) Higher magnification of αD-mCherry and Hbδ-GFP- transfected cells grown under normoxia (above) and hypoxia (below). Scale bar 10 μm. Hbδ formed aggregates and there was more capping in hypoxia-grown cells. (D) Cells transfected with integrin β1-mCherry and Hbδ-GFP showed partial co-distribution under normoxia. The magnification is the same as in .

Article Snippet: The primers for Hbβ, Hbδ, and αD integrin were obtained from Origene.

Techniques: Staining, Transfection

Journal: Life Science Alliance

Article Title: Hypoxia favors tumor growth in colorectal cancer in an integrin αDβ1/hemoglobin δ-dependent manner

doi: 10.26508/lsa.202402925

Figure Lengend Snippet: (A) Blotting of β1 and β2 integrin polypeptides from Colo205 and U937 cell lysates. Colo205 cells contained only β1, whereas U937 cells contained both β1 and β2 integrin chains. (B) Blotting of integrin αD from a slab gel of immune precipitates obtained with control and integrin β1 antibodies (Santa Cruz and BD Biosciences) from Colo205 cells. (C) Immune precipitation with Hb antibody (Thermo Fisher Scientific) from cells grown under normoxia (N) and hypoxia (H), and visualization of integrin αD by blotting the gels with αD antibody. The integrin αD band was obtained from cells grown under hypoxia, but not from cells grown under normoxia. (D) Schematic structure of the αDβ1 integrin bound to the Hbδ tetramer (upper left), and predicted structure of the Hbδ tetramer bound to the integrin αD I-domain (upper right). The possible interacting peptide loops of Hbδ are below. (D, G) The sequence in site 1 is somewhat like the RGD sequence containing the amino acids (D, G) implicated in binding. The other important sequence is quite different. (E) Predicted structure of the Hbδ tetramer bound to the αD I-domain. The possible interacting peptide loops of Hbδ are below. (D, G) The sequence in site 1 is somewhat like the RGD sequence containing the amino acids (D, G) implicated in binding. The other important sequence is quite different. (F) Sequence of Hbδ used for cell transfections. The locations of the I-domain interacting sequences are highlighted in red and green, and the amino acids, which differ between Hbδ and Hbβ are highlighted in blue. (G) Sequence of Hbβ. Source data are available for this figure.

Article Snippet: The primers for Hbβ, Hbδ, and αD integrin were obtained from Origene.

Techniques: Control, Sequencing, Binding Assay, Transfection

Journal: Life Science Alliance

Article Title: Hypoxia favors tumor growth in colorectal cancer in an integrin αDβ1/hemoglobin δ-dependent manner

doi: 10.26508/lsa.202402925

Figure Lengend Snippet: (A) Adhesion of cells to Hb grown under hypoxia and normoxia. Cells grown under hypoxia strongly adhered to red blood cell Hb, whereas those grown under normoxia did not. (B) The αD and β1 integrin antibodies effectively blocked adhesion, whereas the β2 integrin-blocking antibody showed no effect. The RGD-containing peptide also blocked cell adhesion, whereas the control peptide did not. (C) Binding of cells to fibronectin. Soluble Hb (250 μg/ml) and αD antibodies significantly blocked adhesion of cells grown in hypoxia. (D) Oxygen consumption by cells grown under hypoxia was inhibited by the Hbδ/β monoclonal antibody and the Hbδ-derived peptide QLSELHCDKL (400 μM), but not by the corresponding Hbβ-derived peptide ATLSELHCDKL. (E) Proliferation of hypoxia-grown cells was efficiently blocked by RNAi down-regulation of either Hbδ or integrin αD, but not by RNAi of Hbβ. *** P < 0.001, **** P < 0.0001, ** P < 0.01, * P < 0.05.

Article Snippet: The primers for Hbβ, Hbδ, and αD integrin were obtained from Origene.

Techniques: Blocking Assay, Control, Binding Assay, Derivative Assay

Journal: Life Science Alliance

Article Title: Hypoxia favors tumor growth in colorectal cancer in an integrin αDβ1/hemoglobin δ-dependent manner

doi: 10.26508/lsa.202402925

Figure Lengend Snippet: (A) Non-transfected cells bound to red cell Hb. (B) αD-mCherry-transfected cells showed increased binding. Integrin β1 antibodies blocked binding. (C) The cells bound well to recombinant Hbδ, and the adhesion was inhibited by integrin β1 and αD antibodies, and the RGD motif-containing peptide. Average of three parallel results. The experiments were repeated with similar results.

Article Snippet: The primers for Hbβ, Hbδ, and αD integrin were obtained from Origene.

Techniques: Transfection, Binding Assay, Recombinant

Journal: Life Science Alliance

Article Title: Hypoxia favors tumor growth in colorectal cancer in an integrin αDβ1/hemoglobin δ-dependent manner

doi: 10.26508/lsa.202402925

Figure Lengend Snippet: (A) The integrin αD antibody inhibited oxygen consumption, whereas the integrin αX antibody showed no effect. (B) The integrin αD antibody blocked cell proliferation, whereas the integrin αX antibody did not. Average of three parallel measurements. The experiments were repeated with similar results.

Article Snippet: The primers for Hbβ, Hbδ, and αD integrin were obtained from Origene.

Techniques:

Journal: Life Science Alliance

Article Title: Hypoxia favors tumor growth in colorectal cancer in an integrin αDβ1/hemoglobin δ-dependent manner

doi: 10.26508/lsa.202402925

Figure Lengend Snippet: (A) A cancer stained with Hbδ antibody. The tumor tissue is in the middle and lower right corner. (B) An adjacent section from the same tumor stained with mouse IgG. (C) Staining of a cancer with Hbδ antibody. (D) Staining of a consecutive section with Hbα antibody shows no staining of the cancer but positivity of red cells in the vessels. (E) Staining of a cancer with Hbδ antibody shows positive staining of the tumor. (F) Staining of a consecutive section of the same tumor with integrin αD antibody demonstrates co-expression of Hbδ and αD. (G) Staining with integrin β1 antibody. The cancer cells were strongly positive, as were the normal tissue to the right. (H) Staining of with an integrin β2 antibody. The tumor remained negative, whereas the leukocytes in normal tissue were strongly positive. (I) Four images of Hbδ-stained samples in a tissue microarray, magnification 100x. (J) Hbδ staining, magnification 100x. (K) Four examples of αD staining of a tissue microarray, magnification 100x. (L) Integrin αD staining. Original magnification 100x.

Article Snippet: The primers for Hbβ, Hbδ, and αD integrin were obtained from Origene.

Techniques: Staining, Expressing, Microarray

Journal: Life Science Alliance

Article Title: Hypoxia favors tumor growth in colorectal cancer in an integrin αDβ1/hemoglobin δ-dependent manner

doi: 10.26508/lsa.202402925

Figure Lengend Snippet: (A) Whereas the epithelium remains negative, weak staining with anti-Hbδ is found in erythrocytes. (B) Anti-integrin αD stains mainly interstitial macrophages. Original magnification 100x.

Article Snippet: The primers for Hbβ, Hbδ, and αD integrin were obtained from Origene.

Techniques: Staining

Journal: Science Advances

Article Title: Phenotypic discovery and therapeutic evaluation of an ITGA3B1 -targeting antibody-drug conjugate for bladder cancer

doi: 10.1126/sciadv.ady0041

Figure Lengend Snippet: ( A ) ELISA assessing the binding specificity of 2E7 scFv-Fc binding to the ITGA3B1 heterodimer and its individual subunits ( ITGA3 and ITGB1 ), as well as to the structurally related integrin complex ITGA6B4 and its subunits ( ITGA6 and ITGB4 ). HuIgG and BSA were included as negative controls. Data represent the mean ± SD from three technical replicates. A 450 , absorbance at 450 nm. ( B ) Dose-dependent binding of 2E7 scFv-Fc to immobilized ITGA3 , ITGB1 , and ITGA3B1 , as determined by ELISA. ( C ) Flow cytometry analysis of HEK293 cells transiently transfected with ITGA3 , ITGB1 , or both. ( D ) Flow cytometry of MDA-MB-231 cells following siRNA knockdown of ITGA3 , ITGB1 , or both ITGA3 and ITGB1 . ( E ) SPR sensorgrams showing 2E7 scFv-Fc binding to recombinant ITGA3B1 in the presence of either divalent cations or EDTA. Data shown are representative of three independent experiments.

Article Snippet: Recombinant human integrin subunits ( ITGA3 , ITGA6 , ITGB1 , and ITGB4 ; OriGene Technologies, Rockville, MD) and heterodimers ( ITGA3B1 and ITGA6B4 ; Acrobiosystems, Newark, DE) were each coated onto 96-well Maxisorp plates (Thermo Fisher Scientific) at a concentration of 2 μg/ml in PBS and incubated overnight at 4°C.

Techniques: Enzyme-linked Immunosorbent Assay, Binding Assay, Flow Cytometry, Transfection, Knockdown, Recombinant

Journal: Biology

Article Title: Effect of Maternal Hyperglycemia on Fetal Pancreatic Islet Development.

doi: 10.3390/biology14060728

Figure Lengend Snippet: Figure 3. Islet of Langerhans basal membrane protein immunostaining. (A,C,E,G): Immunohis- tochemistry of pan-laminin, laminin alpha 1 and gamma 1 chains and alpha 3 integrin. Islet of Langerhans—dashed line. Brown staining indicates protein-of-interest staining. Scale bar: 20.0 µm. (B,D,F,H): Quantification of stained area related to total islet area for each analyzed protein. Data presented as mean ± standard error of the mean, * p < 0,02 and *** p < 0.0001 vs. control group by Student’s t-test.

Article Snippet: For protein detection, sections were incubated with specific primary rabbit antibodies: pan-laminin (ab1575; Abcam, diluted 1:500, Cambridge, UK), alpha 1 laminin (sc H-300 Santa Cruz H-300: SC-5582 diluted 1:50, Santa Cruz, CA, USA), gamma 1 laminin (sc5584; Santa Cruz, diluted 1:50), alpha 3 integrin (600-401 R13, Rockland diluted 1:200, Baltimore, MD, USA), anti-PCNA (ab2426 Abcam 1: 200), anti-glucagon (# A0565 1:350) and anti-insulin (# A0564 1:250).

Techniques: Membrane, Immunostaining, Staining, Control