Like her pony counterpart, human Twilight is well-spoken and highly intelligent. She holds a great interest in knowledge and science, being able to assemble intricate scientific devices and solve complex mathematical equations, and her intellect rivals that of Sunset Shimmer. She is a bit of a wallflower, acting very awkward and timid around others, especially the standoffish students of Crystal Prep. Despite her problems in socializing, Twilight is eager to connect with people on a personal level, demonstrated in her interactions with Fluttershy, when the two share their love for their pets, and Pinkie Pie, at the Crystal Prep welcoming party. Twilight is shown increasingly distressed by the apparent harm caused by her magic-sensing and storage device to the point where Sunset's eventual accusations leave her distraught.

Applejack is depicted as very reliable, hardworking, and honest. As her friends tend to be quirky, Applejack typically acts as the "straight man" in her circle of friends—much like her Equestrian counterpart tends to be for her friends. She is also considerably strong, able to carry a folding table by herself that normally takes two male athletes to move. Applejack is also the most level-headed among her friends, able to calmly and logically assess situations that she and the others find themselves in. In Rainbow Rocks, she is the first to question the Rainbooms' ability to "pony up", and in Friendship Games, she is the first one to wonder how Rainbow Dash ponies up without playing her guitar.

Like her pony counterpart, Rarity is very generous and has a keen eye for fashion. She disguises Twilight to hide her from the CHS students' ridicule, and she comes up with the idea of handing out pony ears and tails. Rarity also likes doting on the dragon-turned-puppy Spike, finding him "so adorable" and letting him sleep in her lap at the Sweet Shoppe. Rarity's generosity and love for fashion carry over into the second and third films as well. In Rainbow Rocks, she greatly concerns herself with making a good stage impression, and in Friendship Games, she says putting effort into clothes and spending time on my friends are what she lives for.

Pinkie Pie shares many of her pony counterpart's character traits and cartoonish quirks, including having a very cheerful and sociable personality and performing various comedic gags. At one point in the first film, she picks one of the balloon patterns off her skirt and inflates it like an actual balloon. Pinkie displays more cartoonish gags. She has a tendency to skip instead of walk normally, she has a refrigerator stocked with cans of whipped cream, and she tends to pop out of random places such as a locker and air vent. Though she doesn't own a party cannon in the first film, she uses her kick-drums as a party cannon in the second film, and she owns two party cannons by the third film.

Like her pony counterpart, Fluttershy is typically very timid and soft-spoken. She speaks only in barely audible whispers when she first meets Twilight in the first film, and she shrinks away from the spotlight in Rainbow Rocks. Fluttershy has a deep adoration for animals; she instantly opens up to Twilight once she sees Spike for the first time, she regularly sneaks her pets to school in her backpack, and she volunteers at an animal shelter. When Spike reveals that he can talk and is actually a dragon, Fluttershy becomes very excited to meet a talking animal. Fluttershy's strongest trait is her kindness, as displayed in her interactions with animals and various characters. she tries to cheer up human Twilight despite her being from a rivaling school. She is also more confident and self-assured in the second and third films and tie-in animated shorts than she is in the first film.

Like her pony counterpart, Rainbow Dash is very athletic and competitive. She is the captain of every sports team at Canterlot High School, and she effortlessly overtakes Twilight Sparkle in a game of one-on-one. She is also somewhat self-centered, often speaking—and sometimes singing—about her own awesomeness. Rainbow Dash's strongest trait is her loyalty. She eagerly teams up with Twilight to beat Sunset in the first film, and in the third film, she gives up a victory in the Friendship Games to save Sunset's life. She also able to inspire motivation in others as demonstrated in Friendship Games' CHS Rally Song.

Throughout the first film, Sunset Shimmer is shown to be self-absorbed, condescending, manipulative, and dishonest. She frequently insults and belittles others and thinks little of it. According to human Fluttershy, Sunset makes life miserable for "anyone who stands in her way". As a result, most of the Canterlot High School student body is afraid of her. She is also not above making threats and blackmail in order to realize her goals. When Sunset puts on the element of magic, she transforms into a demonic visage of herself. While in this form, she becomes more ruthless, going so far as attempting to dispose of Twilight. After she is defeated, Sunset Shimmer becomes immediately repentant and apologizes for everything she's done, appearing to renounce her prior dishonest and unkind ways.