At the rough endoplasmic reticulum (RER), the initial steps in the creation of integral membrane proteins with multiple transmembrane segments are similar to those seen in amino-end-in proteins.

     The SRP and SRP receptor assist in embedding the first internal signal-anchor sequence. However, further translation results in additional hydrophobic amino acid domains that alternate as stop-transfer anchor and signal-anchor sequences. These insert into the membrane, creating loops in and out of the RER membrane. The SRP and SRP receptors  may not be necessary for the insertion of these additional helices.

     These proteins are eventually transported to the plasma membrane.