Journal: bioRxiv

Article Title: Physiological analysis of the mechanism of Ci transcription factor activation through multiple Fused phosphorylation sites in Hedgehog signal transduction

doi: 10.1101/2025.01.24.634727

Figure Lengend Snippet: ( A-E ) Third instar wing discs with one copy of Ci-WT ( crCi-WT / ci 94 ), showing a stripe of induction of ptc-lacZ reporter expression (“Ptc-lacZ”), visualized by Beta-galactosidase antibody staining (red), in anterior cells near the center of the disc responding to Hh, released from posterior (right) cells. The posterior edge of Ptc-lacZ defines the AP compartment boundary and is marked by yellow dotted lines. ( A ’, B ’, D, E‘ ’) Full-length Ci (Ci-155) visualized by 2A1 antibody staining (gray-scale), and ( C ’) en expression visualized by En antibody staining (green). Anterior En induction by Hh is to the left of the AP border (yellow dotted lines) and co-incides with lower Ci-155 levels, which are otherwise elevated in Hh signaling territory. ( A-C ’) shows wild-type discs using ( A-A ’) 20x and ( B-C ’) 63x objective lenses. ( D-E ’) A Su(fu) LP/LP disc using ( D-D‘ ) 20x and ( E-E’ ) 63x objective lenses, showing normal Ptc-lacZ but lower levels of Ci-155 in anterior and AP border cells. ( F-M ) Third instar wing discs (63x objective) with one copy of the indicated ci CRISPR allele, GFP marking homozygous cos2 mutant clones (green; yellow arrowheads), and yellow dotted lines marking the AP border. ( F ’ -M ’) Ptc-lacZ expression (red) and ( F ’ -M ’’) Ci-155 expression (gray-scale) in the same discs. ( F ’) Ptc-lacZ induction by Ci-WT in cos2 mutant clones was (G ’ ) increased in discs that lack Su(fu) activity, and ( H‘-M‘ ) was increased to a lesser degree by each of the Ci variants shown. ( F” ) Ci-155 levels are increased in cos2 mutant clones because proteolytic processing is blocked, but the increase was reduced or absent ( G” ) in the absence of Su(fu) and ( H”-M” ) for most Ci variants, potentially due to reduced Su(fu)-dependent Ci stabilization or En-mediated inhibition of ci transcription; ( K” ) Ci Δ272-346 and ( L” ) Ci Δ270-300 have higher C-155 than other Ci variants. The brightness of G”-J” and M” images was increased relative to others in order to see any changes in Ci levels in clones more easily. Quantitation used 20x images, all obtained under the same conditions. Scale bars are ( A, D ) 100μm, ( B, C, E ) 20μm, and (all others) 40μm ( N, O ) Bar graphs showing average ( N ) Ptc-lacZ intensity or ( O ) Ci-155 signal intensity in cos2 clones relative to the AP border of control discs (with two copies of Ci-WT), together with SEMs (n values 20, 10, 16, 22, 53, 29, 27, and 31, respectively, for each graph). Ci-WT ratios align with blue dotted lines. Differences with p<0.005 (Student’s t test with Welch correction) are indicated for comparing a Ci variant to ( N ) Ci-WT (black asterisk) or ( O ) Ci Δ1370-1397 (red asterisk). Please see Materials and Methods for details of measurements and expression of all experimental values relative to AP border values of control wild-type wing discs, and Fig1_data supplementary information for raw data, including all p values. A cartoon of Ci domains and deletions, together with testing for En induction in cos2 clones, are shown in Fig. S1.

Article Snippet: For ptc-lacZ protein product staining (“Ptc-lacZ”), rabbit anti-ß-galactosidase (MP Biomedicals) in 1:10,000 dilution was used.

Techniques: Expressing, Staining, CRISPR, Mutagenesis, Clone Assay, Activity Assay, Inhibition, Quantitation Assay, Control, Variant Assay

Journal: bioRxiv

Article Title: Physiological analysis of the mechanism of Ci transcription factor activation through multiple Fused phosphorylation sites in Hedgehog signal transduction

doi: 10.1101/2025.01.24.634727

Figure Lengend Snippet: ( A-H ) Third instar fu mH63 wing discs with one copy of the indicated Ci variant showing ptc-lacZ expression (Ptc-lacZ; red) and ( A ’ -H ’) full-length Ci (Ci-155; gray-scale). ( B-B ’) taken at higher magnification (63x versus 20x objective for others). ( A, A”, B ) Ptc-lacZ is greatly reduced by the loss of Fu kinase activity but expands further anterior in the absence of Fu kinase activity (lower Ptc protein levels allow Hh to travel further); ( A” ) shows the same Ptc-lacZ image shown in ( A ) but zoomed in and in gray-scale with contrast adjusted to show the width of weak expression. ( A ’, B ’) High Ci-155 levels extend further anterior and right up to the AP border (yellow dotted line) in the absence of Fu kinase. ( C-C ’) Ptc-lacZ is increased, with both Ptc-lacZ and Ci-155 stripes narrowed in the additional absence of Su(fu). ( D-H ) Ptc-lacZ was also increased, to a lesser degree, by the tested Ci alterations, together with narrowing of the widths of the elevated Ptc-lacZ and Ci-155 stripes. ( C ’ -H ’) Ci-155 levels were reduced substantially by loss of Su(fu) and for the Ci variants shown, other than for Ci Δ272-346 and Ci Δ270-300. Scale bars are ( B, B ’) 20μm and (all others) 100μm. ( I, J ) Bar graphs showing the average ratio of ( I ) Ptc-lacZ intensity at the AP border of fu mH63 discs relative to the AP border of fu mH63 ; Su(fu) LP/LP discs with one copy of Ci-WT, or ( J ) Ci-155 intensity relative to fu mH63 ; Su(fu) LP/LP wing discs expressing Ci-WT, and then adjusted to the wild-type wing disc control value, together with SEM values (n values 55, 43, 13, 10, 18, 8, and 9, respectively, for each graph). Differences with p<0.005 (Student’s t test with Welch correction) are indicated for comparing a Ci variant to ( I, J ) Ci-WT (black asterisk) or ( J ) Ci Δ1370-1397 (red asterisk). Wing discs for variants with deletions neighboring the SYGHI Su(fu) binding site variably showed enlargement of the anterior compartment. This likely reflects impaired function of the processed repressor derivative, Ci-75, leading to localized de-repression of dpp [ , ]. Please see Materials and Methods for details of measurements and expression of all experimental values relative to AP border values of control wild-type wing discs, and Fig2_data supplementary information for raw data.

Article Snippet: For ptc-lacZ protein product staining (“Ptc-lacZ”), rabbit anti-ß-galactosidase (MP Biomedicals) in 1:10,000 dilution was used.

Techniques: Variant Assay, Expressing, Activity Assay, Control, Binding Assay

Journal: bioRxiv

Article Title: Physiological analysis of the mechanism of Ci transcription factor activation through multiple Fused phosphorylation sites in Hedgehog signal transduction

doi: 10.1101/2025.01.24.634727

Figure Lengend Snippet: ( A-F ) Third instar wing discs (20x objective) with one copy of the indicated ci CRISPR allele, GFP marking homozygous smo GAP-Fu mutant clones (green; yellow arrowheads), and yellow dotted lines marking the AP border. ( A ’ -F ’) Ptc-lacZ (red) and ( A”-F” ) Ci-155 (gray-scale) in the same discs. Fu kinase activity in the clones increased Ptc-lacZ expression and Ci-155 levels in all cases but Ptc-lacZ was induced most strongly by Ci Δ272-346 and Ci Δ270-300. Scale bars are 100μm for all images. ( G, H ) Bar graphs showing the average ratio of ( G ) Ptc-lacZ intensity or ( H ) Ci-155 intensity in smo GAP-Fu clones relative to the AP border of wild-type control discs, together with SEMs (n values 40, 9, 8 15, 19, and 20, respectively, for each graph). Differences with p<0.005 (Student’s t test with Welch correction) are indicated for comparing a Ci variant to ( G, H ) Ci-WT (black asterisk). Please see Materials and Methods for details of measurements and expression of all experimental values relative to AP border values of control wild-type wing discs, and Fig3_data supplementary information for raw data.

Article Snippet: For ptc-lacZ protein product staining (“Ptc-lacZ”), rabbit anti-ß-galactosidase (MP Biomedicals) in 1:10,000 dilution was used.

Techniques: CRISPR, Mutagenesis, Clone Assay, Activity Assay, Expressing, Control, Variant Assay

Journal: bioRxiv

Article Title: Physiological analysis of the mechanism of Ci transcription factor activation through multiple Fused phosphorylation sites in Hedgehog signal transduction

doi: 10.1101/2025.01.24.634727

Figure Lengend Snippet: ( A, B ) Third instar wing disc with one copy of Ci-WT-Sufu (20x objective), showing (A) Ptc-lacZ (red) and ( A ’) Ci-155 expression (gray-scale) or ( B) Ptc-lacZ (red) and ( B ’) En expression (green) at the AP border (yellow dotted line; 63x objective), showing no induction of anterior En. ( C ) Third instar wing disc with one copy of Ci-SYAAD-Sufu (20x objective), showing Ptc-lacZ (red) and ( C ’) Ci-155 expression (gray-scale). ( D-K ) Third instar wing discs (63x objective) with one copy of ( D, F, H, J ) Ci-WT-Sufu or ( E, G, I, K ) Ci-SYAAD-Sufu, GFP marking the indicated clone types (green; yellow arrowheads), and yellow dotted lines marking the AP border. ( D ’ -K ’) Ptc-lacZ expression (red) and ( D”-K” ) Ci-155 expression (gray-scale) in the same discs. ( F, G ) Su(fu) activity was absent in the whole disc. Ptc-lacZ induction in all clones was much lower for both Ci variants than for Ci-WT, but was higher for Ci-SYAAD-Sufu. Scale bars are 100μm for ( A, C ), 20μm for ( B ) and 40μm for ( D-K” ). ( L ) Bar graph showing the average ratio of ( L ) Ptc-lacZ intensity in clones relative to the AP border of wild-type control discs, together with SEMs (n values 20, 5, 14, 10, 87, 19, 40, 54, 46, 47, 199, and 90). Differences with p<0.005 (Student’s t test with Welch correction) are indicated for comparing a Ci variant to Ci-WT (black asterisk) or comparisons between bracketed pairs (red asterisk). Please see Materials and Methods for details of measurements and expression of all experimental values relative to AP border values of control wild-type wing discs, and Fig4 plus S2_data supplementary information for raw data. A bar graph of Ci-155 levels in the clones shown here for Ptc-lacZ, together with examination of pka mutant clones for Ci-WT-Sufu and Ci-SYAAD-Sufu, are shown in Fig. S2.

Article Snippet: For ptc-lacZ protein product staining (“Ptc-lacZ”), rabbit anti-ß-galactosidase (MP Biomedicals) in 1:10,000 dilution was used.

Techniques: Expressing, Activity Assay, Clone Assay, Control, Variant Assay, Mutagenesis

Journal: bioRxiv

Article Title: Physiological analysis of the mechanism of Ci transcription factor activation through multiple Fused phosphorylation sites in Hedgehog signal transduction

doi: 10.1101/2025.01.24.634727

Figure Lengend Snippet: ( A-J ) Third instar wing discs with one copy of each Ci variant showing ( A, C, E, H ) Ptc-lacZ (red) and ( A ’, C ’, E ’, H’ ) Ci-155 expression (gray-scale) (20x objective) or ( B, D, F, I ) Ptc-lacZ (red) and ( B ’, D ’, F ’, I ’) En expression (green) (63X objective), with the AP border marked by dotted yellow lines. Induction of Ptc-lacZ was unchanged for ( A ) Ci-A1 and reduced progressively more by ( C ) Ci-A2, ( E ) Ci-A1A2 and ( H ) Ci ΔΔ (which lacks residues 175-230 and 1201-1271). En induction was normal for ( B ’) Ci-A1, weak for ( D ’) Ci-A2 and absent for ( F ’) Ci-A1A2 and ( I ’) Ci ΔΔ. ( G, J ) fu mH63 discs with one copy of the indicated Ci variant showing Ptc-lacZ (red) and ( G ’, J‘ ) Ci-155 expression (gray-scale). ( K-P ) Third instar wing discs (63x objective) with one copy of the named Ci variant, GFP marking the indicated clone types (green; yellow arrowheads), and yellow dotted lines marking the AP border. ( K ’ -P ’) Ptc-lacZ expression (red) and ( K”-P” ) Ci-155 expression (gray-scale) in the same discs. Ptc-lacZ (red) was not induced by activated Fu kinase for ( K ’) Ci-A1A2 or ( L ’) Ci ΔΔ, even in the absence of cos2 activity (compare O ’ and P ’ to M ’ and N ’; see bar graphs in ). (’’ K -P ’) Ci-155 was elevated similarly for all Ci genotypes by GAP-Fu and by loss of Cos2. Scale bars are 100μM for ( A, C, E, G, H, J ), 20μm for ( B, D, F, I ) and 40 μm for ( K-P ). Quantitative results are included in the graphs of for ( K-P ) and Fig. S5 for ( A-I ). Please see Materials and Methods for details of measurements and expression of all experimental values relative to AP border values of control wild-type wing discs, and Fig6_data and FigS5_data supplementary information for raw data. See Fig. S3 for GAP-Fu clones with Ci-A1 and Ci-A2; Ci Δ1201-1271 (wild-type discs, fu mH63 discs, cos2 and GAP-Fu clones); cos2 GAP-Fu clones (Ci-WT, Ci ΔΔ, Ci Δ175-230, Ci Δ270-300, Ci Δ1370-1397).

Article Snippet: For ptc-lacZ protein product staining (“Ptc-lacZ”), rabbit anti-ß-galactosidase (MP Biomedicals) in 1:10,000 dilution was used.

Techniques: Variant Assay, Expressing, Activity Assay, Control, Clone Assay

Journal: bioRxiv

Article Title: Physiological analysis of the mechanism of Ci transcription factor activation through multiple Fused phosphorylation sites in Hedgehog signal transduction

doi: 10.1101/2025.01.24.634727

Figure Lengend Snippet: ( A, D-I ) Third instar wing discs (63x objective) with ( A, D, E ) one copy of Ci-D1D2 or ( F-I ) the named Ci variant, GFP marking the indicated clone types (green; yellow arrowheads), and yellow dotted lines marking the AP border. ( F-I ) shows smo GAP-Fu clones in Su(fu) mutant discs. ( A ’, D ’ -I ’) Ptc-lacZ expression (red) and ( A”, D”-I” ) Ci-155 expression (gray-scale) in the same discs. ( B ) fu mH63 disc with one copy of Ci-D1D2, showing ( B ) Ptc-lacZ (red) and ( B ’) Ci-155 expression (gray-scale) (20X objective). Wild-type disc with Ci-D1D2, with (C ) Ptc-lacZ (red) and ( C ’) En expression (green) at the AP border (yellow dotted line; 63X objective). Scale bars are 100μm for ( B ), 20μm for ( C ) and 40μm for all other images. ( J, L ) Bar graphs showing the ratio of Ptc-lacZ intensity in smo GAP-Fu clones in ( J ) otherwise wild-type or ( L ) Su(fu) mutant discs relative to the AP border of control discs, together with SEMs (n= 40, 31, 113, 10, 72, 28, 15, 25, 120, 55, and 14 respectively in ( J ) and 79, 70, 45, 30, 75, 99, and 35, respectively, in ( L )). ( K ) Bar graph showing the ratio of Ptc-lacZ intensity in cos2 clones (pink) and in smo cos2 GAP-Fu clones (red) relative to the AP border of control discs, together with SEMs (n= 20, 38, 45, 67, 38, 55, 30, 22, 53, and 36 respectively for cos2 and 47, 49, 7, 112, 75, 104, 15, 74, and 38, respectively, for smo cos2 GAP-Fu ). ( J-L ) Differences with p<0.005 (Student’s t test with Welch correction) are indicated for comparing a Ci variant to Ci-WT (black asterisk) or comparisons between bracketed pairs (red asterisk). Grey dotted brackets compare cos2 clones and blue dotted brackets compare cos2 GAP-Fu clones for Ci-D1D2. Please see Materials and Methods for details of measurements and expression of all experimental values relative to AP border values of control wild-type wing discs, and Fig6_data supplementary information for raw data.

Article Snippet: For ptc-lacZ protein product staining (“Ptc-lacZ”), rabbit anti-ß-galactosidase (MP Biomedicals) in 1:10,000 dilution was used.

Techniques: Variant Assay, Clone Assay, Mutagenesis, Expressing, Control

Journal: bioRxiv

Article Title: Physiological analysis of the mechanism of Ci transcription factor activation through multiple Fused phosphorylation sites in Hedgehog signal transduction

doi: 10.1101/2025.01.24.634727

Figure Lengend Snippet: ( A-F, I, J, R-T ) One copy of named Ci variants in ( A, B, E, F, I, J, Q, R ) otherwise wild-type, ( C, D ) Su(fu) LP/LP , or ( G, H, S ) fu mH63 third instar wing discs, showing ( A-J, Q-S ) Ptc-lacZ (red), ( A ’, C ’, E‘, G ’, I ’, Q ’, S ’) Ci-155 (gray-scale) or ( B ’, D ’, F ’, H ’, J ’, R ’) En expression (green, with the AP border marked by dotted yellow lines. ( K-P ) Third instar wing discs (63x objective) with the named Ci variant, GFP marking the indicated clone types (green; yellow arrowheads), and yellow dotted lines marking the AP border. ( K ’ -P ’) Ptc-lacZ expression (red) and ( K”-P” ) Ci-155 expression (gray-scale) in the same discs. ( L, M, N, P ) GAP-Fu and cos2 GAP-Fu clones also lack smo activity, and ( N ) GAP-Fu; Su(fu ) indicates lack of Su(fu) activity in the whole disc. Scale bars are 100μm for ( A, G, I, Q, S ), 20μm for ( B-F, H, J, R ) and 40μm for all other images. Quantitative results are included in the graphs of for all clones shown and Fig. S5 for the AP border of wild-type and fu mH63 discs.

Article Snippet: For ptc-lacZ protein product staining (“Ptc-lacZ”), rabbit anti-ß-galactosidase (MP Biomedicals) in 1:10,000 dilution was used.

Techniques: Expressing, Variant Assay, Clone Assay, Activity Assay

Journal: bioRxiv

Article Title: Physiological analysis of the mechanism of Ci transcription factor activation through multiple Fused phosphorylation sites in Hedgehog signal transduction

doi: 10.1101/2025.01.24.634727

Figure Lengend Snippet: ( A-F ) Third instar wing discs with one copy of the named Ci variant (63X objective), showing GFP expression (green) to mark smo cos2 GAP-Fu clones (yellow arrowheads) and ptc-lacZ (red) to mark the AP border in wing discs with one genomic copy of a transgene encoding ( A, C, E ) wild-type Cos2 (gCos-WT) or ( B, D, F ) Cos2 S572A S931A (gCos-AA). ( A ’ -F‘ ) Ci-155 expression (gray-scale); AP border marked by yellow dotted lines on the basis of Ptc-lacZ expression (not shown). Ci-155 was clearly higher in all clones with gCos-WT but was more similar to surrounding anterior cells away from the AP border in the presence of gCos-AA, with the clearest distinction observed for ( F ’) Ci-A1A2. Scale bars are 40μm for all images. ( G ) Bar graph showing the ratio of Ci-155 intensity in smo cos2 GAP-Fu clones relative to the AP border of wild-type control discs, together with SEMs (n= 116, 52, 21, and 96, respectively for gCos-WT and 76, 38, 26, and 98, respectively, for gCos-AA) for the designated Ci variants together with gCos-WT (gray) or gCos-AA (black). ( H ) Bar graph showing the difference in the ratio of Ci-155 intensity between smo cos2 GAP-Fu clones and the surrounding anterior cells, relative to the AP border of wild-type control discs, together with SEMs (n values the same as G ) for designated Ci variants together with gCos-WT (gray) or gCos-AA (black). ( G ) Differences with p<0.005 (Student’s t test with Welch correction) are indicated for differences between values for gCos-WT versus gCos-AA for a given Ci (red asterisk). ( H ) Differences with p<0.005 (Student’s t test with Welch correction) are indicated for comparing the change in Ci-155 level induced by the clone genotype relative to zero (red asterisk). Please see Materials and Methods for details of measurements and expression of all experimental values relative to AP border values of control wild-type wing discs, and Fig7_data supplementary information for raw data. See Fig. S5 for the effect of loss of Fu sites in Su(fu) and Cos2 on the activity of Ci-A1A2.

Article Snippet: For ptc-lacZ protein product staining (“Ptc-lacZ”), rabbit anti-ß-galactosidase (MP Biomedicals) in 1:10,000 dilution was used.

Techniques: Variant Assay, Expressing, Clone Assay, Control, Activity Assay

Journal: Cell Death & Disease

Article Title: Targeting CK2 eliminates senescent cells and prolongs lifespan in Zmpste24 -deficient mice

doi: 10.1038/s41419-024-06760-0

Figure Lengend Snippet: A A Pie chart indicating the different functional groups of drugs in the DDR-related kinase inhibitors library used in the screen. B The primary screen of all 80 DDR-related kinase inhibitors at 10 μM concentration. X-axis – number of senescent cells in the drug-treated cultures relative to cells treated with vehicle only; Y-axis – fraction of total cells remaining after drug treatment relative to vehicle-treated controls; gray shaded area shows drugs that reduce the proportion of senescent cells by > 70%. C Independent validation of the primary screen expressed as cell senescence and cell number relative to untreated control cultures of senescent cells. All drugs were used at 10 μM, chart plots mean ± SD from triplicate experiments. * p < 0.05, ** p < 0.01 by two-tailed Student’s t test. D Representative images derived from triplicate experiments of passage 6 Zmpste24 −/− MEF cultures measuring senescence-associated β-galactosidase (SA-β-Gal) activity using X-gal staining. Scale bar, 50 μm. E Quantification of SA-β-Gal–positive staining from 10 randomly chosen fields of view for each group. The chart plots mean ± SEM. * p < 0.05, ** p < 0.01.

Article Snippet: The chemical library were purchased from Targetmol Chemicals Inc. Cellular Senescence Detection Kit was purchased from Cell Signaling Technology (#9860).

Techniques: Functional Assay, Concentration Assay, Biomarker Discovery, Control, Two Tailed Test, Derivative Assay, Activity Assay, Staining

Journal: Cell Death & Disease

Article Title: Targeting CK2 eliminates senescent cells and prolongs lifespan in Zmpste24 -deficient mice

doi: 10.1038/s41419-024-06760-0

Figure Lengend Snippet: A Senescence-associated β-galactosidase staining in Ck2α or scramble siRNA treated Zmpste24 +/+ and Zmpste24 −/− MEFs at passage 6 (scale bar, 200 μm). B Representative immunoblots showing the levels of proCaspase-3 (ProCasp-3), cleaved caspase-3 (cCasp-3) and P16 in Ck2α or scramble siRNA-treated Zmpste24 −/− MEFs and wild-type controls. C Representative flow cytometric plots for assessing apoptosis in Ck2α or scramble siRNA-treated Zmpste24 −/− MEFs and wild-type controls. D Quantitation of the percentage of viable (gate II: PI − /annexin V − ) and apoptotic (gates III and IV: PI − /annexin V + and PI + /annexin V + ) cells in ( C ); chart plots mean ± SD from independent triplicate experiments. E Representative immunoblots showing protein levels of γH2AX at indicated time points after 4 μM camptothecin (CPT) for 1 h to induce DNA damage or vehicle DMSO control in siRNA-Ck2α transfected in Zmpste24 −/− MEFs and wild-type controls treated with 75 μM TBB for 6 h at the indicated time (after CPT treatment 0, 2, 4 h). At least three independent experiments were performed.

Article Snippet: The chemical library were purchased from Targetmol Chemicals Inc. Cellular Senescence Detection Kit was purchased from Cell Signaling Technology (#9860).

Techniques: Staining, Western Blot, Quantitation Assay, Control, Transfection

Journal: Cell Death & Disease

Article Title: Targeting CK2 eliminates senescent cells and prolongs lifespan in Zmpste24 -deficient mice

doi: 10.1038/s41419-024-06760-0

Figure Lengend Snippet: In normal cells undergoing DNA damage, CK2α regulates heterochromatin relaxation by phosphorylating HP1α, which promotes its dynamic dissociation with H3K9me3 and relocalization at DNA double-strand break sites (DSB), which facilitates DNA damage response and repair. Reducing CK2 activity leads to the accumulation of DNA damage, which leads to cellular senescence. Since CK2 activity is low in Zmpste24 -deficient cells (progeria cells), CK2 inhibition by TBB aggravates extant DDR defects in senescent cells and results in apoptosis. CK2 activity determines the fate of cells undergoing varying degrees of DNA damage.

Article Snippet: The chemical library were purchased from Targetmol Chemicals Inc. Cellular Senescence Detection Kit was purchased from Cell Signaling Technology (#9860).

Techniques: Activity Assay, Inhibition