Prominent conservative Twitter users pushed back hard on Wednesday after an overnight purge of their accounts that culled thousands of their followers, as the embattled social media platform faced mounting charges of left-wing bias and suppression of right-wing viewpoints.

San Francisco-based Twitter said it acted to reduce the flow of fake news and abusive content by eliminating accounts suspected of being Russian-created “bots,” but conservative Twitter users, including podcast host Dan Bongino and Michael Flynn Jr., son of former National Security Adviser Michael Flynn, said the company is effectively targeting their right to free speech.

“The Twitter purge is real,” Mr. Bongino said on his Twitter account.

Conservative Project Veritas investigative filmmaker James O’Keefe commented sarcastically on his Twitter account Wednesday: “Looks like thousands of Twitter users committed the thought crime of tweeting about ‘God,’ ‘the American flag’ and ‘guns,’ and were taken off the platform.”

As the hashtag #TwitterLockOut trended across the internet, the first of what is likely to be multiple lawsuits was filed against Twitter. Jared Taylor, founder and editor of the white nationalist magazine American Renaissance, announced that he was suing Twitter for viewpoint discrimination.

Mr. Taylor said his account was suspended in December as part of a Twitter crackdown against users affiliated with hate groups, in reaction to the use of social media during the deadly white nationalist rally in August in Charlottesville, Virginia.

The firm claimed it was banning “specific threats of violence or wish for the serious physical harm, death, or disease of an individual or group of people.”

Mr. Taylor denied that his group promoted violence in any way. His attorney, Noah Peters, made the novel legal argument this week that Twitter violated a California law protecting free speech in public spaces.

That law, legal analysts say, had not previously been applied to social media or the internet.

“Our lawsuit is not about whether Taylor is right or wrong,” Mr. Peters wrote online. “It’s about whether Twitter and other technology companies have the right to ban individuals from using their services based on their perceived viewpoints and affiliations.”

He said “Twitter censorship” should terrify everyone.

“Allowing Twitter to censor content is extremely troublesome given Twitter’s self-proclaimed mission to ‘give everyone the power to create and share ideas instantly, without barriers.’”

The battle between the so-called alt-right movement and Silicon Valley over a perceived bias against conservatives stems back to at least 2015, when conservative activist Charles Johnson filed a lawsuit after he was banned from Twitter. Conservatives have also contended that Facebook has a bias in its algorithms and in its “curators” to suppress right-wing news content.

Twitter has yet to say how many accounts it has purged this week and has not commented on the Jared Taylor lawsuit. But it has repeatedly insisted that its social media platform is politically neutral.

“Twitter’s tools are apolitical, and we enforce our rules without political bias,” the company said in a statement. “As part of our ongoing work in safety, we identify suspicious account behaviors that indicate automated activity or violations of our policies around having multiple accounts, or abuse.”

On Thursday, the firm faced more pressure to explain its methodologies in the wake of special counsel Robert Mueller’s indictment last week of 13 Russian nationals and entities on charges of meddling in the 2016 U.S. presidential election. The indictment detailed how Russian operatives used fake Twitter accounts to conduct “information warfare” against the U.S. political system.

Much of the issue revolves around “bots” — accounts powered by automated software. Researchers estimate that as many as 15 percent of all Twitter accounts could be fake or controlled by bots.

The Kremlin-linked “troll farm” cited in Mr. Mueller’s indictment used bots to exaggerate U.S. political divisions on hot-button cultural and political issues, helping these issues and ideas rise to the top of the trending metrics.

During Capitol Hill hearings into Russian election meddling last year, lawmakers lashed out at Twitter — along with internet giants Facebook and Google — for not confronting the Russian bot problem earlier and more seriously.

Last month, in response to reporting by The New York Times, Twitter closed more than 1 million fake accounts.

Military analysts continually express deep concern over Russia’s ability to manipulate bots to ratchet up anger across American social media and deepen partisan divisions.

Last year, the bipartisan Alliance for Securing Democracy created a program to track Kremlin-backed Twitter accounts. The Alliance’s Hamilton 68 project watches for pulses from Moscow that appear to amplify corrosive cultural debates in America.

Late Thursday, #TwitterLockOut was the leading hashtag being promoted by accounts linked to the Kremlin.

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