The CHP said the incident occurred in the eastbound lanes of the freeway near 150th Avenue, after 4 p.m. A short time later, a victim with a gunshot wound showed up at Eden Medical Center in Castro Valley.
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The shooting marks the second on that stretch of road in a matter of weeks.
Thursday afternoon, ABC7 News captured video of two cars riddled with bullet holes, being towed from Eden Medical Center. However, the CHP said the suspect vehicle remains outstanding.
The agency explained it does not believe the second vehicle was involved in the incident on the freeway, but the vehicle became part of the investigation as officers began their follow-up.
SKY7 captured the scene and the traffic backup that followed cars stretched for miles.
The incident comes one day after an East Bay couple decided to share their chilling story and shocking video. Footage recorded two weeks ago shows Garrett Mason and fiancée Tina Do in the middle of a shootout as a vehicle cut them off and another tailed them.
VIDEO: Bay Area couple opens up about surviving I-580 shooting as bullet 'went right past' their faces
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Couple opens up about I-580 shooting as bullet shoots through car
The pair were leaving work, and similar to Thursday's incident, they were heading eastbound on I-580 in San Leandro.
The city's mayor Pauline Russo Cutter addressed the violence.
"It's got to be a two-pronged approach. We've got to attack it as it is right now and go after the people who are doing this now," Cutter said. "But we also have to bring generations up to not have this kind of gun violence. And quite frankly, I believe that we need to do much more than we're doing right now at the federal level with gun control."
CHP said the motive for this incident is under investigation. It is now added to a list of more than 80 freeway shootings in the Bay Area in the last four years.
"Unfortunately, we were in the wrong place at the wrong time. Also, luck at the right time," Mason and Do told ABC7 News. "People have commented, 'If you were a second faster in your car or a second slower, how could that have changed where the bullet went?' But we can't think too much about the what-ifs. It's just where we're at."
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RELATED: CHP reveals Alameda Co. had 76 freeway shootings in past year
76 freeway shootings in Alameda Co. in last 12 months, CHP says
Dozens of drivers and passengers on Bay Area freeways have been hit by bullets sometimes caught in the crossfire, other times targeted.
"It's not something that we should tolerate," Mayor Cutter told ABC7 News. "I mean, we can't tolerate this."
Anyone with information or who may have witnessed the incident is asked to contact the CHP investigative Tip Line at (707)917-4491