this is a colloquial phrase that we use to emphasize the previous point.
here's an example:
A: we were really busy at work today
B: yeah, I know.
A: no, seriously. I didn't even get to take any breaks
so in this example, although person B agrees that it was busy, person A adds "no seriously" to emphasize just how busy it was.
*it doesn't really matter whether person B agrees or disagrees with person A, it can be used any time you want to add extra emphasis on the previous statement.
example:
A:This class is so hard!
B: I don't think it's that bad
A: No, seriously! I took chemistry last semester and it wasn't as hard as this class.