Haley's comments come at a United Nations Security Council meeting that centered on Iran after the US requested an emergency session amid protests in the country.
"Human rights are not the gift of governments, they are the inalienable rights of the people themselves," Haley said. "The Iranian regime is now on notice, the world will be watching what you do."
She added: "Freedom and human dignity cannot be separated from peace and security. When the rights of the people are denied, the people rightly resist. If the concerns are not acknowledged, then peace and security are inevitably threatened."
Haley said that "in the end, the Iranian people will determine their own destiny," and she reiterated US support for the protesters.
"And let there be no doubt whatsoever, the United States stands unapologetically with those in Iran who seek freedom for themselves, prosperity for their families and dignity for their nation," she added.
While most nations echoed Haley's support for human rights and free speech, some also voiced concern that the UN Security Council was not the appropriate venue to discuss protests in Iran.
"It is up to the Iranians, and to the Iranians alone, to pursue the path of peaceful dialogue, a dialogue based on full respect for the fundamental rights and freedoms of the Iranian people," French Ambassador Francois Delattre said. "However worrying the events of the last few days in Iran may be, they do not constitute, per se, a threat to international peace and security."
Prior to the meeting, Delattre warned against the "instrumentalisation of the crisis from the outside because it would only reinforce the extremes, which is precisely what we want to avoid."
Russian UN Ambassador Vassily Nebenzia also slammed the US for using the meeting to bring up the Iranian protests under a "bogus pretext."
"Let Iran deal with its own problems," said Nebenzia, who once again raised the idea of the Security Council meeting about protests in Ferguson, Missouri, and Occupy Wall Street demonstrations as the Russian mission to the UN did in a tweet on Tuesday.
A representative from Iran responded forcefully at Friday's meeting.
"The move by the United States to bring to this council protests in Iran by some of our citizens for their legitimate grievances -- some exacerbated by none other than the US itself in its dereliction of its obligations under the Joint Comprehensive Plan of Action -- is an abuse of its power as a permanent member, and an abuse of the council itself," Iranian Ambassador to the UN Gholamali Khoshroo said.
"It is unfortunate that despite the resistance on the part of some of its members, this council has allowed itself to be abused by the current US administration in holding a meeting on an issue that falls outside the scope of its mandate, putting on display the failure of the council to fulfill its real responsibility in maintaining international peace and security," he said.
The US has been vocal in its support of the demonstrators and offered harsh condemnation of the Iranian government, including a tweet from President Donald Trump that called the Iranian regime "brutal and corrupt."
For its part, Iran has accused the US of "grotesque" meddling in social media to incite unrest -- and thus tampering with Iranian affairs, according to a letter sent to the UN on Thursday.
Haley said earlier this week that the US wanted emergency meetings in New York and in Geneva at the Human Rights Council to discuss the Iran situation.
Russia's deputy foreign minister has called the US proposal "harmful and destructive."
Differences in opinion between the US and Europe over how to address the situation in Iran began to manifest themselves ahead of Friday's meeting.
"The United States has spoken clearly and unequivocally," Vice President Mike Pence wrote in a Washington Post
op-ed Wednesday. "Unfortunately, many of our European partners, as well as the United Nations, have thus far failed to forcefully speak out on the growing crisis in Iran. It's time for them to stand up."
While the US and its European partners -- including the United Kingdom, France and Germany -- all fundamentally support the right of Iranian demonstrators to peacefully protest, they disagree in their analysis of where the situation on the ground is headed and over the strategic value of ramping up the rhetoric publicly, sources have told CNN.
Uncertainty over how long the protests will continue has prompted European countries to collectively take a much more cautious approach to what it says publicly than the US.