The motion was defeated by 122 votes to 97, Deputy Speaker Pollachi V Jayaraman, who presided over the proceedings, said.
Earlier, the Deputy Speaker went for voice vote twice and declared that the motion had been defeated but the DMK insisted on a division which Jayaraman permitted.
Subsequently, 97 MLAs -- members of the DMK and allies Congress and IUML -- voted in favour of the motion moved by Opposition leader M K Stalin, while 122 ruling party MLAs voted against it.
Those who voted against the motion included Mylapore MLA R Nataraj who had voted against the Edappadi K Palaniswami government during the February 18 floor test.
The MLAs belonging to the O Panneerselvam faction in the AIADMK including the former chief minister himself were not present during the voting.
After the division was taken and the votes counted, Jayaraman declared that 97 MLAs had voted in favour of the motion and 122 against it.
"The motion is defeated," he said, amid cheering and thumping of desks by ruling party members.
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