We recently reported that a precisely positioned in-tether chiral center can modulate backbone peptides' secondary structures, which provides an unbiased platform to evaluate peptides' biophysical properties solely imposed by secondary structure differences. In this work, we studied the cellular uptake efficiency and mechanism of epimer pairs of a panel of chirality-induced helical peptides (CIH peptides). Although the peptides' cellular uptake is a synergetic result of various factors, our results unambiguously indicate that helical content is an important factor for the cellular uptake of CIH peptides.