Journal: Cell death discovery

Article Title: Loss of TC-PTP in keratinocytes leads to increased UVB-induced autophagy.

doi: 10.1038/s41420-025-02353-8

Figure Lengend Snippet: Fig. 2 Generation and characterization of HaCaT TC-PTP knockout (KO) keratinocytes. TC-PTP/Mock (engineered control) and HaCaT TC- PTP/KO cells were generated using the CRISPR/Cas9 genome editing system. A Representative photomicrographs of HaCaT TC-PTP/KO and TC-PTP/Mock cells after 3 days of culture. All three clones selected for both cell lines were cultured under the same conditions. B Immunoblot analysis of HaCaT TC-PTP/Mock and TC-PTP/KO cell lysates with antibodies specific for TC-PTP, CD44, pSTAT3, STAT3, and β-actin. A431 epidermoid carcinoma cell line was used as a positive control. C–F Cell proliferation of the selected clones (C1–C3) of HaCaT TC-PTP/Mock and TC-PTP/KO cells. Proliferation of the cells was measured using WST-8 assay according to the manufacturer’s general manual. The results are the mean ± standard deviation from three independent experiments. *P < 0.005 by T-test for equality of means. C HaCaT clones C1. D HaCaT clones C2. E HaCaT clones C3. F HaCaT clones C1–C3.

Article Snippet: To generate HaCaT TC-PTP/KO cell lines, HaCaT cells were transfected with either the CRISPR/Cas9 plasmid targeting TC-PTP (sc-403071, Santa Cruz Biotechnology) or the corresponding control plasmid (sc-418922, Santa Cruz Biotechnology).

Techniques: Knock-Out, Control, Generated, CRISPR, Clone Assay, Cell Culture, Western Blot, Positive Control, Standard Deviation

Journal: Cell death discovery

Article Title: Loss of TC-PTP in keratinocytes leads to increased UVB-induced autophagy.

doi: 10.1038/s41420-025-02353-8

Figure Lengend Snippet: Fig. 3 TC-PTP deficiency decreases apoptosis and increases autophagy in human HaCaT keratinocytes following UVB exposure. A, B HaCaT TC-PTP/Mock and TC-PTP/KO cells were exposed to 5, 10, or 20 mJ/cm2 of UVB irradiation and incubated for 16 h following UVB exposure. A Representative photomicrographs of HaCaT TC-PTP/Mock and TC-PTP/KO cells after UVB exposure. Scale bar: 100 μm. B Quantitative analysis of the percentage of apoptotic cells characterized by cell ballooning, nuclear condensation, and bleb formation. After 16 h of UVB treatment, apoptotic keratinocytes were counted microscopically in at least three non-overlapping fields. Results are the mean ± standard deviation from three independent experiments. *P < 0.05 by T-test for equality of means. C, D HaCaT TC-PTP/Mock and TC- PTP/KO cells exposed to 10 mJ/cm2 of UVB irradiation and cells were harvested after 16 h following UVB exposure. C Immunoblot analysis of HaCaT TC-PTP/Mock and TC-PTP/KO cell lysates with antibodies specific for LC3, cleaved PARP, cleaved caspase-3, TC-PTP, and β-actin. D Immunoblot analysis of HaCaT TC-PTP/Mock and TC-PTP/KO cell lysates with antibodies specific for LC3, Bcl-2, Bax, TC-PTP, and β-actin. E Cell viability was measured using WST-8 assay. *P < 0.005 by T-test for equality of means. F Immunoblot analysis of HaCaT TC-PTP/Mock and TC- PTP/KO cell lysates with antibodies specific for LC3, SQSTM1, TC-PTP, and β-actin. TC-PTP/Mock and TC-PTP/KO cells were exposed to 10 mJ/cm2 of UVB and harvested at the indicated time post-UV irradiation. Total cell lysates were then prepared.

Article Snippet: To generate HaCaT TC-PTP/KO cell lines, HaCaT cells were transfected with either the CRISPR/Cas9 plasmid targeting TC-PTP (sc-403071, Santa Cruz Biotechnology) or the corresponding control plasmid (sc-418922, Santa Cruz Biotechnology).

Techniques: Irradiation, Incubation, Standard Deviation, Western Blot

Journal: Cell death discovery

Article Title: Loss of TC-PTP in keratinocytes leads to increased UVB-induced autophagy.

doi: 10.1038/s41420-025-02353-8

Figure Lengend Snippet: Fig. 4 Inhibition of autophagy in keratinocyte survival and proliferation following UVB exposure. A, B Effect of inhibition of autophagy on keratinocyte survival and proliferation in response to UVB irradiation. TC-PTP/Mock and TC-PTP/KO cells were pretreated with 3-MA (5 mM) for 1-h prior exposure to UV irradiation (10 mJ/cm2). Cells were then collected 16 h after UVB irradiation. A Immunoblot analysis of HaCaT TC-PTP/ Mock and TC-PTP/KO cell lysates with antibodies specific for LC3, TC-PTP, and β-actin. B Cell viability of the cells was measured using WST-8 assay. *P < 0.005 by T-test for equality of means. C, D Effect of inhibition of autophagy on keratinocyte survival and proliferation in response to UVB irradiation. TC-PTP/Mock and TC-PTP/KO cells were pretreated with CQ (50 μM) for 1-h prior exposure to UV irradiation (10 mJ/cm2). Cells were then collected 16 h after UVB irradiation. C Immunoblot analysis of HaCaT TC-PTP/Mock and TC-PTP/KO cell lysates with antibodies specific for LC3, TC-PTP, and β-actin. D Cell viability of the cells was measured using WST-8 assay. *P < 0.005 by T-test for equality of means.

Article Snippet: To generate HaCaT TC-PTP/KO cell lines, HaCaT cells were transfected with either the CRISPR/Cas9 plasmid targeting TC-PTP (sc-403071, Santa Cruz Biotechnology) or the corresponding control plasmid (sc-418922, Santa Cruz Biotechnology).

Techniques: Inhibition, Irradiation, Western Blot

Journal: Cell death discovery

Article Title: Loss of TC-PTP in keratinocytes leads to increased UVB-induced autophagy.

doi: 10.1038/s41420-025-02353-8

Figure Lengend Snippet: Fig. 5 Inhibition of autophagy on the regulation of UVB-induced apoptosis in keratinocytes. A–C TC-PTP/Mock and TC-PTP/KO cells were pretreated with 3-MA (5 mM) for 1-h prior exposure to UV irradiation (10 mJ/cm2). Cells were then collected 16 h after UVB irradiation. A Representative photomicrographs of HaCaT TC-PTP/Mock and TC-PTP/KO cells after UVB exposure in the presence or absence of 3-MA. Scale bar: 100 μm. B Quantitative analysis of the percentage of apoptotic cells characterized by cell ballooning, nuclear condensation, and bleb formation. After 16 h of UVB treatment, apoptotic keratinocytes were counted microscopically in at least three non-overlapping fields. Results are the mean ± standard deviation from three independent experiments. *P < 0.05 by T-test for equality of means. C, D TC-PTP/Mock and TC-PTP/KO cells were pretreated with CQ (50 μM) for 1-h prior exposure to UV irradiation (10 mJ/cm2). Cells were then collected 16 h after UVB irradiation. C Representative photomicrographs of HaCaT TC-PTP/Mock and TC-PTP/KO cells after UVB exposure in the presence or absence of CQ. Scale bar: 100 μm. D Quantitative analysis of the percentage of apoptotic cells characterized by cell ballooning, nuclear condensation, and bleb formation. After 16 h of UVB treatment, apoptotic keratinocytes were counted microscopically in at least three non- overlapping fields. Results are the mean ± standard deviation from three independent experiments. *P < 0.05 by T-test for equality of means. E, F TC-PTP/Mock and TC-PTP/KO cells were pretreated with CQ (50 μM) for 1-h prior to exposure to UV irradiation (10 mJ/cm2). Apoptotic cells were stained with Annexin V-FITC and estimated using flow cytometry analysis. E Representative outputs of flow cytometry analysis. F Quantification of apoptotic cells in control, UVB-treated, and CQ-UVB-treated HaCaT TC-PTP/Mock and KO cells 16 h post-UVB irradiation. The results are the mean ± standard deviation from three independent experiments. *P < 0.05 by T-test for equality of means.

Article Snippet: To generate HaCaT TC-PTP/KO cell lines, HaCaT cells were transfected with either the CRISPR/Cas9 plasmid targeting TC-PTP (sc-403071, Santa Cruz Biotechnology) or the corresponding control plasmid (sc-418922, Santa Cruz Biotechnology).

Techniques: Inhibition, Irradiation, Standard Deviation, Staining, Cytometry, Control