Figure 7
From: hMMS2 serves a redundant role in human PCNA polyubiquitination
MMS2 null mouse emybronic stem show no overt disruption of PCNA polyubiquitination after UV irradiation. (A) RT-PCR for MMS2 was performed as described in material and methods (B) Wild type and MMS2 null embryonic stem cells were lysed and subjected to immunoblotting with anti-Mms2/Uev1a antibody. (C) Confluent plates of wild type and MMS2 null embryonic stem cells were split 1:4 on Day 1. On Day 2 media was replaced and followed by UV irradiation with either 0 or 30 J/m2 on Day 3. Cells were harvested 6 hours post-treatment and subjected to immunoblotting with an anti-PCNA antibody. (D) Wild type and MMS2 null embryonic stem cells were transfected with Uev1a and lysed 72 hours later followed by immunoblotting with anti-Mms2/Uev1a antibody. (E) MMS2-/- cells were transfected with siRNA targeting mouse UEV1a. 72 hours post transfection cells were UV irradiated, lysed and subjected to immunoblotting using an anti-PCNA antibody.