anti phosphorylated chk1  (Cell Signaling Technology Inc)

Journal: Archives of biochemistry and biophysics

Article Title: Pericentriolar material 1 aggregation maintains cell survival upon prolonged replication stress.

doi: 10.1016/j.abb.2025.110383

Figure Lengend Snippet: Fig. 7. PCM1 promotes CHK1 activation to maintain cell survival upon prolonged replication stress. (A) Depletion of PCM1 did not affect CHK2 activation upon HU treatment. Extracts of control or PCM1-depleted cells were analyzed by western blotting with antibodies against phosphorylated CHK2, CHK2, and tubulin (loading control). (B) Inhibition of CHK2 did not affect U2-OS cell viability upon HU treatment. Quantitative results of relative cell numbers of HU-treated U2-OS cells in the presence of CHK2 inhibitor (CHK2i). (C) Depletion of PCM1 reduced CHK1 activation upon HU treatment. Extracts of control or PCM1-depleted cells were analyzed by western blotting with antibodies against phosphorylated CHK1, CHK1, and tubulin (loading control). (B) Inhibition of CHK1 reduced U2-OS cell viability upon HU treatment. Quantitative results of relative cell numbers of HU-treated U2-OS cells in the presence of CHK1 inhibitor (CHK1i). n.s.: no significance, ***: P < 0.001. Each experiment was repeated at least three times.

Article Snippet: The following antibodies were used for immunofluorescence and Western blot assays: anti-γ-tubulin (Sigma), anti-PCM1, anti-phosphorylated ATR, anti-ATR, anti-CHK1, anti-CHK2, anti-phosphorylated CHK1, anti-phosphorylated CHK2, anti-LC3 A/B (Cell Signaling), antip150glued, anti-GM130 (BD Biosciences), anti-Ku70, anti-α-tubulin, antiATM, anti-β-actin (GeneTex), anti-γ-H2AX, anti-p-ATM (Abcam), antiphosphorylated DNA-PKcs, anti-DNA-PKcs (Santa Cruz Biotech).

Techniques: Activation Assay, Control, Western Blot, Inhibition

Journal: Archives of biochemistry and biophysics

Article Title: Pericentriolar material 1 aggregation maintains cell survival upon prolonged replication stress.

doi: 10.1016/j.abb.2025.110383

Figure Lengend Snippet: Fig. 8. Autophagy maintains U2-OS cell viability upon prolonged replication stress. (A–B) Autophagy was induced by HU treatment in U2-OS cells. (A) LC3 puncta were examined by immunofluorescence staining with antibodies against LC3. DNA was stained with DAPI. Scale bar: 10 μm. (B) Quantitative results of (A). (C, upper panel) Extracts of HU-treated cells were analyzed by western blotting with antibodies against LC3 and actin. (C, lower panel) Quantitative results of upper panel. (D) Depletion of PCM1 alleviated HU-induced autophagy. Extracts of wild-type or PCM1-depleted (siPCM1) cells in the presence or absence of HU were analyzed by western blotting with antibodies against LC3 and actin. (E) Quantitative results of (D). (F) Inhibition of autophagy reduced U2-OS cell viability upon HU treatment. Quantitative results of relative cell numbers in wild-type or PCM1-depleted (siPCM1) cells in the presence or absence of HU. (G) Graphic abstract of this study. Prolonged replication stress promotes centrosome amplification independent of PCM1 aggregation. However, PCM1 promotes DNA damage signaling the ATM-CHK1 axis and autophagy to sustain cell survival. n.s.: no significance, ***: P < 0.001. Each experiment was repeated at least three times.

Article Snippet: The following antibodies were used for immunofluorescence and Western blot assays: anti-γ-tubulin (Sigma), anti-PCM1, anti-phosphorylated ATR, anti-ATR, anti-CHK1, anti-CHK2, anti-phosphorylated CHK1, anti-phosphorylated CHK2, anti-LC3 A/B (Cell Signaling), antip150glued, anti-GM130 (BD Biosciences), anti-Ku70, anti-α-tubulin, antiATM, anti-β-actin (GeneTex), anti-γ-H2AX, anti-p-ATM (Abcam), antiphosphorylated DNA-PKcs, anti-DNA-PKcs (Santa Cruz Biotech).

Techniques: Immunofluorescence, Staining, Western Blot, Inhibition, Amplification

Journal: Journal of Cancer

Article Title: Regulation of response to radiotherapy by β-arrestin1 in Non-small cell lung cancer

doi: 10.7150/jca.30012

Figure Lengend Snippet: β-arrestin1 mediates radiation-induced signaling pathway. A. Phosphorylation of ATR and Chk1 were parallel to β-arrestin1 status, especially at 0 and 10 hs after radiation. B,C,D. radiation could activated ERK and NF-kB pathways within 60 minutes.

Article Snippet: Anti-β-arrestin1, -ATR, -Chk1, -H2AX, -ERK, -PARP, and -NF-kB; anti-phosphorylated ATR (Ser428), Chk1 (Ser345), BRCA1 (Ser1524), ERK (Thr202/Tyr204), γ-H2AX (Ser139), and -NF-kB p65 (Ser536) antibodies were all purchased from Cell Signaling Technology, USA.

Techniques: