https://orcid.org/0000-0002-8909-5207
Figures & data
Figure 1. PTPN21 was successfully overexpressed in Jurkat, NALM-6, and Reh cells. (a) Western blot data show that Flag-PTPN21 was overexpressed in Jurkat, NALM-6, and Reh cells. GAPDH was used as the internal control. (b) IF results reveal that Flag-PTPN21 could be detected in the transfected Jurkat, NALM-6, and Reh cells.
Figure 2. Excessive PTPN21 had no effects on apoptosis of Jurkat, NALM-6, or Reh cells. (a) Annexin V/7-AAD assay results reveal that overexpression of PTPN21 did not affect apoptosis of ALL cells. (b) Quantification of the Annexin V/7-AAD results. N.S., no significance.
Figure 3. Overexpression of PTPN21 facilitated proliferation and cell cycle progression in EGF-stimulated Jurkat, NALM-6, and Reh cells. (a) CCK-8 staining results show that overexpression of PTPN21 elevated cell proliferation in all three ALL cell types with EGF-stimulation. (b) Ki-67/7-AAD assay data uncover that the cell cycle progression is promoted by excessive PTPN21 in all three ALL cell lines. (c) Quantification of the Annexin V/7-AAD results. N.S., no significance. *P < 0.05, **P < 0.01, ***P < 0.001.
Figure 4. Overexpression of PTPN21 activated Src and the MAPK signaling pathways in EGF-stimulated ALL cells. (a) Western blot data show that in Jurkat cells, PTPN21 overexpression reduced p-Src level and upregulated p-ERK1/2 and p-JNK levels, but had no impact on p-p38 level. (b) Western blot data reveal that in NALM-6 cells, PTPN21 overexpression led to a decreased level of p-Src and elevated levels of p-ERK1/2 and p-p38 but didn’t affect p-p38 level. (c) Western blot results uncover that in Reh cells, excessive PTPN21 downregulated p-Src level and elevated levels of p-ERK1/2, p-p38, and p-p38.
Figure 5. JNK inhibitor mitigated the pro-proliferative effect of PTPN21 on EGF-stimulated Jurkat cells. (a) CCK-8 staining results reveal that MEK inhibitor PD90859 and JNK inhibitor SP600125 have little impact on PTPN21’s pro-proliferative function in EGF-stimulated Jurkat cells. (b) Ki-67/7-AAD assay data show that the cell cycle progression promoted by excessive PTPN21 in Jurkat cells was not affected by PD90859 but was mitigated by SP600125. (b) Quantification of the Annexin V/7-AAD results. N.S., no significance. *P < 0.05, **P < 0.01, ***P < 0.001.
Figure 6. p38 pathway mediated the pro-proliferative function of PTPN21 in EGF-stimulated NALM-6 cells. (a and b) CCK-8 staining (a) and Ki-67/7-AAD assay data (b) show that the proliferation and cell cycle progression facilitated by PTPN21 overexpression in NALM-6 cells was not influenced by MEK inhibitor PD90859 but was affected by p38 inhibitor SB203580. (c) Quantification of the Annexin V/7-AAD results. N.S., no significance. *P < 0.05, **P < 0.01, ***P < 0.001.
Figure 7. Inhibition of p38 partially attenuated the proliferation of EGF-stimulated Reh cells driven by PTPN21 overexpression. (a) CCK-8 staining assay data show that none of the inhibitors can abolish the proliferation-promoting effect of PTPN21 on Reh cells. (b) Ki-67/7-AAD assay results illustrate that MEK inhibitor PD90859 and JNK inhibitor SP600125 had minimal impact on the cell cycle of EGF-stimulated Reh cells while p38 inhibitor SB203580 significantly increased the proportion of Reh cells at the G1 phase with PTPN21 overexpression. (c) Quantification of the Annexin V/7-AAD results. N.S., no significant difference. *P < 0.05, **P < 0.01, ***P < 0.001.
Data availability statement
The datasets used and/or analyzed during the current study are available from the corresponding author on reasonable request.