5-Star bases much of its appeal on fighting corruption, its flagship policy is a "citizens wage" to help Italy's army of unemployed, and it promises a referendum on membership of the euro currency blamed by many for years of economic underperformance.
Italy's 5Star Movement, the populist challenger to the country's established parties, suffered a shocking defeat on Sunday, lowering the risk of snap elections being called in 2017, according to analysts.
Observers are closely watching Genoa, the hometown of 5-Star Movement leader Beppe Grillo.
It is a stark reversal of fortunes for the populist party and reduces its chances of leading the national government after parliamentary elections next year.
With most ballots counted from elections a day earlier in a thousand small cities and towns, the Five-Star Movement imploded in all big races including in Genoa, home of leader-comic Beppe Grillo.
A closer contest is expected in the northern port city of Genoa, where the center-right hopes to win control from the center-left incumbent.
Newspaper headlines and pundits pounced on the results, saying the 5-Star might have peaked, but Grillo shot back on Monday, confidently predicting his group would bounce back.
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In all but one of the top cities up for grabs, candidates from centre-right and centre-left alliances earned berths in June 25 run-offs, since none clinched more than 50% of the votes.
Sunday's balloting will also test whether the Democratic Party, led by former Premier Matteo Renzi, can rebound locally after suffering stinging losses to the 5-Stars in Rome and Turin mayoral races a year ago.
The center-right, dominated by the Northern League and Silvio Berlusconi's Forza Italia party, is favorite in Verona in the northeast, while the center-left is expected to keep control of L'Aquila, capital of the central Abruzzo region.
The only outright victor in a major city on Sunday was Leoluca Orlando, who led the centre-left to victory in the Sicilian capital Palermo, securing his fifth mandate.
Hence, the most likely scenario continued to be that general elections would wait until the end of the first quarter of 2018 at the earliest.
Until last week, Italy's main parties had been trying to pass a new proportional electoral law together, but the deal unraveled.