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Barcelona's municipal police force said 750,000 people participated in the march on Saturday, called to demand the release of eight former members of the dissolved Catalan government and the presidents of the two main grassroots independence organisations - the Catalan National Assembly (ANC) and Omnium Cultural, reports Efe news.
The ANC and Omnium had organised Saturday's demonstration.
The jailed separatist leaders include former vice president of the Catalan regional government, Oriol Junqueras, who like the rest of his fellow Cabinet members is charged with rebellion, sedition and misuse of public funds for his role in enabling the region's October 27 declaration of independence.
The demonstrators, many waving pro-independence Estelada flags, marched past the Sagrada Familia church and other emblematic parts of Barcelona behind banners that read "Freedom for Political Prisoners" and "We Are a Republic".
Barcelona Mayor Ada Colau also participated .
The former speaker of Catalonia's Parliament, Carme Forcadell, who also is under investigation for rebellion, sedition and misuse of public funds, did not attend Saturday's march on the advice of her attorney.
On Friday, she was released from custody after posting a 150,000 euro ($174,000) bond.
Puigdemont, who fled to Brussels late last month, delivered a recorded message on Friday to the pro-independence movement.
He called on people to take to the streets and said the Spanish government's crackdown on independence leaders would not dissuade the region from freely deciding its future at the ballot box.
Puigdemont is scheduled to appear before a judge in a November 17 hearing on the European arrest warrants that have been issued for him and four officials of his administration who accompanied him to Belgium.
Hours after Catalonia's October 27 declaration of independence, the Spanish Senate approved Prime Minister Mariano Rajoy's proposal to invoke Article 155 of the constitution and rescind Catalonia's regional autonomy.
Rajoy dismissed Puigdemont and his Cabinet, dissolved the Catalan Parliament and called regional elections for December 21