The crash occurred on westbound I-70 near the Silver Lake Rest area in Madison County at about 1:54 a.m., police said.
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Investigators said the Greyhound bus was traveling westbound when it struck three commercial vehicles parked on the exit ramp to the Silver Lake I-70 rest area.
Police said three people in the bus were killed and 14 transported to hospitals, No one inside the commercial vehicles was injured, police said.
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Troopers closed westbound I-70 to traffic between mileposts 28 and 24 as they investigate the crash.
A Greyhound spokesperson released a statement saying, "We can confirm an incident occurred this morning involving Greyhound schedule number 1675 traveling from Indianapolis to St. Louis. Multiple passengers, including our driver, have been transported to the hospital. Our primary concern is ensuring we care for our passengers and driver at this time. We are working closely with local authorities and a relief bus is on the way for passengers."