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. 2022 Dec 12;13:7677. doi: 10.1038/s41467-022-35469-x

Fig. 4. USP22 inhibits ubiquitin-proteasomal degradation of SPI1.

Fig. 4

a, b SPI1 protein expression in PRDM1-overexpressing and vector Hep3B cells (a) or in PRDM1-knockout and sgCtrl Huh7 cells (b). n = 3 independent biological replicates. c SPI1 protein expression was determined in PRDM1 knockout and sgCtrl Huh7 cells. n = 3 independent biological replicates. d SPI1 was pulled down and ubiquitin-conjugated SPI1 was then determined by immunoblotting using an anti-HA antibody. n = 3 independent biological replicates. e SPI1 was pulled down and ubiquitin-conjugated SPI1 was determined by immunoblotting using an anti-HA antibody. n = 3 independent biological replicates. f DUB siRNA library screening indicated that USP22 and USP33 suppression decreased SPI1 protein levels. g qRT-PCR of USP22 or USP33 expression in PRDM1-overexpressing and vector Hep3B cells or in PRDM1-knockout and sgCtrl Huh7 cells. Data presented as mean ± SEM (n = 3 independent biological replicates). h Western blotting of USP22 or USP33 expression in PRDM1-overexpressing and vector Hep3B cells or in PRDM1-knockout and sgCtrl Huh7 cells. n = 3 independent biological replicates. i, j SPI1 protein expression in USP22-overexpressing and vector Hep3B cells (i) or in USP22-knockdown and control Huh7 cells (j). n = 3 independent biological replicates. k SPI1 protein expression was determined in USP22-knockdown or control Huh7 cells. n = 3 independent biological replicates. l SPI1 was pulled down and ubiquitin-conjugated SPI1 was then determined by immunoblotting using an anti-HA antibody. n = 3 independent biological replicates. m SPI1 was pulled down and ubiquitin-conjugated SPI1 was then determined by immunoblotting using an anti-HA antibody. n = 3 independent biological replicates. P value was determined by unpaired two-sided Student’s t test (g). Source data are provided as a Source data file.