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Whether AWS API calls go via public internet or not is determined by your VPC networking setup - specifically VPC Endpoints. For the S3 service you can have either a Gateway or Interface VPC Endpoint. Generally a Gateway one is the way to go unless you need access to the endpoint from on-prem. So if your VPC is set up with an S3 Gateway VPC Endpoint and you have the appropriate route in your route tables, then any AWS API calls to S3 from your EC2 instance will stay on the AWS network.
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