Fig. 3: USP34 is preferentially expressed in GSCs to promote GSC maintenance and GBM tumor growth. | Nature Communications

Fig. 3: USP34 is preferentially expressed in GSCs to promote GSC maintenance and GBM tumor growth.

From: Stabilization of Pin1 by USP34 promotes Ubc9 isomerization and protein sumoylation in glioma stem cells

Fig. 3

a Immunoblot analysis of USP34 in GSCs and NSTCs. b, c Representative images (b) and statistical quantification (c) of immunofluorescent analysis of USP34 (red) and SOX2 or OLIG2 (green) in human primary GBMs. Scale bar, 20 μm. (n = 5 biological replicates; mean ± s.d.) d Immunoblot analysis of USP34 in T4121 GSCs expressing shUSP34. e Representative images of tumorsphere formation of T4121 GSCs expressing shUSP34 or shNT. Forty-eight hours after lentiviral infection, 2000 cells were planted in each well of 96-well plates and cultured for 5 days. Scale bar, 50 μm. f, g Statistical quantifications of tumorsphere numbers (f) and sizes (g) of T4121 GSCs expressing shUSP34 or shNT. Three random 20× fields were used for calculation. (n = 3 biological replicates; ***P < 0.001; mean ± s.d.; two tailed unpaired t-test). h Cell proliferation of T4121 GSCs expressing shUSP34 or shNT. Forty-eight hours after lentiviral infection, 2000 cells were planted in each well of 96-well plates and cell viability was determined at the indicated time points. (n = 4 biologically independent experiments; ***P < 0.001; mean ± s.d.; two-way ANOVA). i In vivo bioluminescent imaging analysis of intracranial growth of GBM xenografts derived from T4121 GSCs expressing shUSP34 or shNT. Five-thousand GSCs were injected into mouse brains and representative images at the indicated days post-transplantation were shown. j Kaplan-Meier survival curves of tumor-bearing mice in (i). (shNT, n = 5 mice; shUSP34#1, n = 7 mice; shUSP34#2, n = 6 mice; shUSP34#1 vs. shNT, P = 0.0018; shUSP34#2 vs. shNT, P = 0.0038; **P < 0.01; two-tailed log-rank test.) k, l Immunofluorescent analysis (k) and statistical quantifications (l) of Ki67 (green) and cleaved caspase-3 (red) in GBM xenografts. Scale bar, 40 μm (n = 5 tumors for each group; ***P < 0.001; mean ± s.d.; two-tailed unpaired t-test). The blotting experiments were repeated at least three times with biological replicates (a, d). Source data are provided as a Source Data file.

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