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Grant Wood's best known painting, American Gothic, is one of the few images to reach the status of universally recognised cultural icon.

I had thought that I should change the bold part to "painting American Gothic" (so effectively removing the commas around the title), since otherwise it isn't exactly clear which painting the author is referring to (yes I know American Gothic is famous, but there may be other paintings that could be referred to). However, apparently the answer is to keep things as they are (so keep the commas), and I was wondering why. Could someone help me understand this?

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This is called a "non-restrictive clause" in English. Using the commas shows that "best known painting" and "American Gothic" refer to the same thing. Dropping the commas here makes the sentence ungrammatical, because Wood only has one best known painting, but you could say this:

Grant Wood's painting American Gothic is one of the most recognizable images in American art.

Here, "American Gothic" is a restrictive clause. "Grant Wood's painting" could refer to any of his paintings; the following clause restricts it to a single one. Restrictive clauses in English generally don't use commas.

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