American Society of Hematology

Figure 3

$Figure 3. cANGPTL4 interacts with integrin α5β1, VE-cadherin, and claudin-5 in a bimodal manner. (A) In situ kinetic PLA detection of various complexes between cANGPTL4 and indicated binding partners in HMVECs treated with 6 μg/mL cANGPTL4. Values (means ± SD) represent mean fold change in the number of interactions compared with the zero time point, as determined from n = 3 independent experiments (600 HMVECs) using BlobFinder. Experiments were terminated when microscopic lesions were observed at 180 minutes after cANGPTL4 treatment. (B) Immunodetection of integrin α5β1, VE-cadherin, and claudin-5 in anti-cANGPTL4 immunoprecipitates from total protein lysates of HMVECs treated with 6 μg/mL rh-cANGPTL4. (C) Immunodetection of integrins β1, integrin α5, integrin β3, VE-cadherin, and claudin-5 from total protein lysate versus internalized fraction of HMVECs treated with cANGPTL4. Protein lysates were collected every 10 minutes (0-180 minutes). Values below each band represent the mean fold change in protein expression level compared with the cognate zero time point (n = 3). *P < .05; **P < .01.$

cANGPTL4 interacts with integrin α5β1, VE-cadherin, and claudin-5 in a bimodal manner. (A) In situ kinetic PLA detection of various complexes between cANGPTL4 and indicated binding partners in HMVECs treated with 6 μg/mL cANGPTL4. Values (means ± SD) represent mean fold change in the number of interactions compared with the zero time point, as determined from n = 3 independent experiments (600 HMVECs) using BlobFinder. Experiments were terminated when microscopic lesions were observed at 180 minutes after cANGPTL4 treatment. (B) Immunodetection of integrin α5β1, VE-cadherin, and claudin-5 in anti-cANGPTL4 immunoprecipitates from total protein lysates of HMVECs treated with 6 μg/mL rh-cANGPTL4. (C) Immunodetection of integrins β1, integrin α5, integrin β3, VE-cadherin, and claudin-5 from total protein lysate versus internalized fraction of HMVECs treated with cANGPTL4. Protein lysates were collected every 10 minutes (0-180 minutes). Values below each band represent the mean fold change in protein expression level compared with the cognate zero time point (n = 3). *P < .05; **P < .01.

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