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. Author manuscript; available in PMC: 2022 Sep 27.
Published in final edited form as: Cell Rep. 2022 Sep 13;40(11):111348. doi: 10.1016/j.celrep.2022.111348

Figure 2. SerpinB3 regulates known CSC pathways.

Figure 2.

(A) RNA was isolated after SerpinB3 knockdown in T4121 cells, and qPCR was performed for SERPINB3, OCT4, NANOG, OLIG2, MYC, and TGF-β1 (3 technical replicates).

(B) Tumor cells were plated in a limiting-dilution manner, and the number of wells containing spheres was counted after 14 days and used to calculate stem cell frequencies using the online algorithm detailed in the methods. N = 12 wells per dilution.

(C) c-MYC expression after SerpinB3 knockdown was assessed via western blot, with actin as a loading control.

(D) TGF-β1 secretion was analyzed 2 days after plating and normalized to total protein (3 technical replicates per condition, per tumor model).

(E) NanoString pathway score comparing SerpinB3 knockdown to non-target control. Bolded rows represent pathways known to regulate the CSC state.

(F) Immunofluorescence staining of T4121 CSCs for SerpinB3 and SOX2. Scale bar represents 10 μM.

p < 0.05 was considered statistically significant. *p < 0.05, **p < 0.01, ***p < 0.001, as determined by 1-way ANOVA with Dunnett’s multiple comparisons, Student’s t test for qPCR data, or chi-squared p value for limiting dilution assay. Error bars represent standard deviations.