The United States had on Friday agreed to withdraw its more than 1,000 soldiers from the African nation where Washington built a $100 million base to fly a fleet of drones.
The demonstration in the northern desert town of Agadez, home to a US air base, was called by a group of 24 civil society associations that have backed the regime since last year's coup.
"This is Agadez, not Washington, US army go home," read a big banner held up by protesters.
Issouf Emoud, who fronts the M62 Movement in the town, told AFP: "Our message is clear: American soldiers pack your bags and go home".
He had also organised demonstrations demanding the departure of French forces which pulled out last year.
Niger has long been a linchpin in the US and French strategy to combat jihadists in West Africa.
The Nigerien military announced last month it was breaking off a defence agreement with the United States, claiming it had been imposed and the US troop presence was illegal.
US Deputy Secretary of State Kurt Campbell agreed to remove the troops in a meeting in Washington with Niamey's prime minister, Ali Mahaman Lamine Zeine, US officials told AFP on condition of anonymity.
The US military deployment in Niger "is of no use for our security", said civil society leader Amobi Arandishu.
"The armed groups still rage across the desert," he told AFP. "Russians, Americans, Germans, French, they all come here for their own interests."
Following the overthrow of elected president Mohamed Bazoum last July, the junta kicked out troops from former colonial power France before the end of 2023.
Russian military instructors arrived in Niger this month with an air defence system and other equipment, state media said, after talks between military ruler General Abdourahamane Tiani and Russian President Vladimir Putin.
The US pullout marks a new regional gain for Russia, which has ramped up its focus on Africa, backing military regimes in neighbouring Mali and Burkina Faso.
Niger faces violence by Boko Haram jihadists and their rivals Islamic State West Africa Province (ISWAP) from the southeastern region of Diffa near Nigeria.
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