Western blot analysis Nuclear and mitochondrial protein was extracted with RIPA buffer (89901, ThermoFisher Scientific) supplemented with 1% Protease Inhibitor Cocktail Set I (539131, Merck, Burlington, MA)
While were on the subject, we dont even like the term rolling retests. You should never try to retest without pulling over at your first opportunity
Firstly, exposure to caregivers who smoked was dichotomized (yes vs no) for each of the three caregiver smoking categories ( primary caregiver smoked , secondary caregiver smoked , and both caregivers smoked ) at Wave 1 and 2 and repeated for each teenage substance use variable at Wave 3 ( teenager ever-smoking , current smoking , ever e-cigarette use , ever alcohol use , current alcohol use , and ever use of other drugs )
Because smoking and nicotine exposure can influence multiple hormone-regulating systems, understanding their potential effects on testosterone provides useful context when evaluating long-term health risks
