The group blocked off the southbound side of the freeway. Protesters then gathered on freeway lanes and unfurled signs, including a large banner that read, "Joe Biden, don't ignore us." The freeway reopened just before 11:30 p.m.
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Azerbaijan's launch of reportedly intense artillery firing in the Nagorno-Karabakh region on Tuesday raised fears that another full-scale conflict with Armenia could be underway, less than three years after a war that killed more than 6,000 people.
A similar demonstration occurred in August in which several hundred protesters blocked one side of the 134 Freeway in Glendale to call attention to the crisis in Artsakh.
READ MORE | Protesters block 134 Freeway in Glendale, creating massive traffic backup
Protesters block 134 Freeway in Glendale, creating massive backup
On Wednesday, Azerbaijan and Armenian forces reached a cease-fire agreement to end the two days of fighting the separatist region that has been a flashpoint for decades, officials on both sides said.
An hour after the truce was announced, Armenian Prime Minister Nikol Pashinyan said that the intensity of the hostilities in the region "has decreased drastically." Azerbaijani authorities said they had halted the military operation launched a day earlier once separatist officials said they were laying down arms.
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It wasn't immediately clear if some fighting continued, but both sides agreed to talks Thursday on the "re-integration" of the region into Azerbaijan. That, in addition to guarantees to lay down arms, was widely viewed as a victory for Baku.
The Associated Press contributed to this report.