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Enhanced ubiquitination and phosphorylation in UVB-treated coelomocytes was confirmed using western blot. Representative western blots showing (a) ubiquitinated, (c) phosphorylated ATM/ATR substrate, (e) phosphorylated AKT substrate, (g) <t>phosphorylated</t> <t>MAPK/CDK</t> substrate, and (i) phosphorylated <t>PKC</t> substrate signal in biological triplicate controls (C, white font) and UVB-treated (U, red font) samples. Beta actin loading controls are shown below. Quantification and statistical testing (one-tailed paired t-test) of western blot signals are shown in (b) , (d) , (f) , (h) , and (j) . Individual data points representing n = 6 biological replicates are overlaid as white circles. Bars represent mean values, and error bars represent ± the standard deviation of n = 6 biological replicates. NS, not significant (p > 0.05). * p ≤ 0.05, ** p ≤ 0.01. P-MAPK/CDK and P-PKC blots were run simultaneously and therefore have the same β actin loading control. Additional western blots are presented in <xref ref-type=Supplementary Figure 4 . " width="250" height="auto" />
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Enhanced ubiquitination and phosphorylation in UVB-treated coelomocytes was confirmed using western blot. Representative western blots showing (a) ubiquitinated, (c) phosphorylated ATM/ATR substrate, (e) phosphorylated AKT substrate, (g) <t>phosphorylated</t> <t>MAPK/CDK</t> substrate, and (i) phosphorylated <t>PKC</t> substrate signal in biological triplicate controls (C, white font) and UVB-treated (U, red font) samples. Beta actin loading controls are shown below. Quantification and statistical testing (one-tailed paired t-test) of western blot signals are shown in (b) , (d) , (f) , (h) , and (j) . Individual data points representing n = 6 biological replicates are overlaid as white circles. Bars represent mean values, and error bars represent ± the standard deviation of n = 6 biological replicates. NS, not significant (p > 0.05). * p ≤ 0.05, ** p ≤ 0.01. P-MAPK/CDK and P-PKC blots were run simultaneously and therefore have the same β actin loading control. Additional western blots are presented in <xref ref-type=Supplementary Figure 4 . " width="250" height="auto" />
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Enhanced ubiquitination and phosphorylation in UVB-treated coelomocytes was confirmed using western blot. Representative western blots showing (a) ubiquitinated, (c) phosphorylated ATM/ATR substrate, (e) phosphorylated AKT substrate, (g) <t>phosphorylated</t> <t>MAPK/CDK</t> substrate, and (i) phosphorylated <t>PKC</t> substrate signal in biological triplicate controls (C, white font) and UVB-treated (U, red font) samples. Beta actin loading controls are shown below. Quantification and statistical testing (one-tailed paired t-test) of western blot signals are shown in (b) , (d) , (f) , (h) , and (j) . Individual data points representing n = 6 biological replicates are overlaid as white circles. Bars represent mean values, and error bars represent ± the standard deviation of n = 6 biological replicates. NS, not significant (p > 0.05). * p ≤ 0.05, ** p ≤ 0.01. P-MAPK/CDK and P-PKC blots were run simultaneously and therefore have the same β actin loading control. Additional western blots are presented in <xref ref-type=Supplementary Figure 4 . " width="250" height="auto" />
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Enhanced ubiquitination and phosphorylation in UVB-treated coelomocytes was confirmed using western blot. Representative western blots showing (a) ubiquitinated, (c) phosphorylated ATM/ATR substrate, (e) phosphorylated AKT substrate, (g) phosphorylated MAPK/CDK substrate, and (i) phosphorylated PKC substrate signal in biological triplicate controls (C, white font) and UVB-treated (U, red font) samples. Beta actin loading controls are shown below. Quantification and statistical testing (one-tailed paired t-test) of western blot signals are shown in (b) , (d) , (f) , (h) , and (j) . Individual data points representing n = 6 biological replicates are overlaid as white circles. Bars represent mean values, and error bars represent ± the standard deviation of n = 6 biological replicates. NS, not significant (p > 0.05). * p ≤ 0.05, ** p ≤ 0.01. P-MAPK/CDK and P-PKC blots were run simultaneously and therefore have the same β actin loading control. Additional western blots are presented in <xref ref-type=Supplementary Figure 4 . " width="100%" height="100%">

Journal: Frontiers in Immunology

Article Title: UVB-induced genotoxic stress activates the DNA damage response and innate immune pathways in sea urchin coelomocytes

doi: 10.3389/fimmu.2026.1787989

Figure Lengend Snippet: Enhanced ubiquitination and phosphorylation in UVB-treated coelomocytes was confirmed using western blot. Representative western blots showing (a) ubiquitinated, (c) phosphorylated ATM/ATR substrate, (e) phosphorylated AKT substrate, (g) phosphorylated MAPK/CDK substrate, and (i) phosphorylated PKC substrate signal in biological triplicate controls (C, white font) and UVB-treated (U, red font) samples. Beta actin loading controls are shown below. Quantification and statistical testing (one-tailed paired t-test) of western blot signals are shown in (b) , (d) , (f) , (h) , and (j) . Individual data points representing n = 6 biological replicates are overlaid as white circles. Bars represent mean values, and error bars represent ± the standard deviation of n = 6 biological replicates. NS, not significant (p > 0.05). * p ≤ 0.05, ** p ≤ 0.01. P-MAPK/CDK and P-PKC blots were run simultaneously and therefore have the same β actin loading control. Additional western blots are presented in Supplementary Figure 4 .

Article Snippet: Primary antibodies included ubiquitin rabbit polyclonal antibody (Cell Signaling Technology, #58395), phospho-ATM/ATR Substrate Motif (Cell Signaling Technology, #6966), phospho-Akt Substrate Motif (Cell Signaling Technology, mix of #9614 and #10001), phospho-MAPK/CDK Substrate Motif (Cell Signaling Technology, mix of #9477 and #2325), and phospho-PKC Substrate Motif (Cell Signaling Technology, #6967). β-actin rabbit polyclonal antibody was used as a loading control (Cell Signaling Technology, #4967, diluted 1:1000 in 1x TBST with 5% w/v BSA).

Techniques: Ubiquitin Proteomics, Phospho-proteomics, Western Blot, One-tailed Test, Standard Deviation, Control