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. 2023 Jul 10;8(13):e157491. doi: 10.1172/jci.insight.157491

Figure 4. Syx regulates the expression of cyclins and cyclin-dependent kinase inhibitors.

Figure 4

(A–D) Immunoblotting (A and C) and qPCR (B and D) analyses of the expression of p21 (CDKN1A), p27 (CDKN1B), Cyclin D1, Cyclin E2 (CCNE2), Cyclin A2 (CCNA2), and Cyclin B1 (CCNB1) in U251 (A and B) and LN229 (C and D) cells expressing indicated shRNAs, grown at subconfluency. For Western blots (A and C), different biological samples are separated by dashed lines. Samples from each blot set were run in parallel, except Cyclin A2 and the corresponding GAPDH from the same gel blot. The same samples for the Cyclin D1 blot with equal loading amounts as other Cyclins blots were run at different times (C, indicated by larger white space). See Figure 2B for the GAPDH loading control for p21 and p27 blots in C. Bar graphs (B and D) represent mean ± SEM of 3–4 biological replicates of relative mRNA expression of indicated genes normalized by GAPDH or β-actin. One-way ANOVA with Dunnett’s multiple-comparison test. *P < 0.05, **P < 0.01, ***P < 0.001.