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. 2019 Aug 5;132(15):1823–1832. doi: 10.1097/CM9.0000000000000294

Figure 4.

Figure 4

Detection of ERS- and apoptosis-related proteins in the mus COL4A3 knockout mouse monoclonal cell line. (A) Schematic diagram of the specific position of each COL4A3 mutation site in the COL4A3 protein. (B) Sequence analysis showed the generation of a knockout COL4A3 c.4852 insA p.Arg1618ThrfsX4 genotype in mouse podocytes using CRISPR/Cas9 technology. (C) Western blotting detection of mouse podocyte mutants (COL4A3 c.4852 insA p.Arg1618ThrfsX4); Clone 1 showed no obvious COL4A3 protein expression. Basal detectable COL4A3 protein expression could be observed in the WT group. This truncated mutation group presented increased ERS-related proteins (GRP78, GRP94, PERK, p-eIF2α, and CHOP), but eIF2α was unchanged. Apoptosis-related protein expression (CHOP and cleaved caspase-3) was upregulated, but Bcl-2 was downregulated. Lane 1 and 2 were duplicates; similar results were observed for the same mutation in clone 2. ERS: Endoplasmic reticulum stress; PERK: PKR-like endoplasmic reticulum kinase; p-eIF2α: Phosphorylated eukaryotic translation initiation factor 2α; WT: Wild-type.