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Fig. 3 | Cell & Bioscience

Fig. 3

From: PSMD2 contributes to the progression of esophageal squamous cell carcinoma by repressing autophagy

Fig. 3

PSMD2 inhibits autophagy by inducing ASS1. A Differential expression analysis of proteomics data of KYSE30 cells with or without PSMD2 knockdown. B Functional enrichment of the differentially expressed proteins in KYSE30 cells with or without PSMD2 knockdown by KEGG. C Functional enrichment of the differentially expressed proteins in KYSE30 cells with or without PSMD2 knockdown by Metascape. D Statistical analysis of the protein levels of of ASS1 detected by IHC of tissue microarray consisting of 144 ESCC and normal samples. E Box plots showing ASS1 expression levels in the early and advanced stage of ESCC. Data were derived from tissue microarray. F The positive correlation between the protein levels of PSMD2 and ASS1 proteins in ESCC tissues. N = 144. G-H KYSE450 cells with PSMD2 overexpression were examined for PSMD2 mRNA levels (G) and ASS1 mRNA levels (H) by qPCR.I, J KYSE450 cells with PSMD2 knockdown were examined for PSMD2 mRNA levels (I) and ASS1 mRNA levels (J) by qPCR.K KYSE30 cells with PSMD2 overexpression or vector were treated with CHX (100 μg/ml). The protein level of ASS1 was examined at 0, 2, 4, 8 h by immunoblotting (left panel) and protein half-life calculated (right panel).L, M ASS1 overexpression reduced the formation of LC3B puncta. Autophagy was induced in KYSE30 cells (L) and KYSE450 cells (M) with ASS1 overexpression by FBS deprivation. The cells were then stained with the DAPGreen (green). The nucleus was stained with Hoechst 33342 (blue). Scale bar: 30 μm. Quantitative analysis of the autophagic puncta was shown in the right panel. N = 3. Data were presented as mean ± SD. N–O ASS1 knockdown in KYSE30 cells (N) and KYSE450 cells (O) increased the formation of LC3B puncta. N = 3. Data were presented as mean ± SD

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