lpv lopinavir  (Ambeed Inc)

Journal: Advanced Science

Article Title: The Compromised Fanconi Anemia Pathway in Prelamin A‐Expressing Cells Contributes to Replication Stress‐Induced Genomic Instability

doi: 10.1002/advs.202307751

Figure Lengend Snippet: Replication stress response defects and genomic instability in prelamin A‐expressing cells. a) SA‐β‐gal staining of IMR90‐hTERT fibroblasts treated with DMSO or LPV (20 µ m ) for 2 weeks. Scale bars: 100 µm. The percentage of SA‐β‐gal‐positive cells was calculated. n = 3. b) The level of γH2AX in prelamin A‐expressing ( ZMPSTE24 ‐knockdown or LPV‐treated) cells was measured by western blotting. n = 3. c) Immunofluorescence staining of 53BP1 in control and prelamin A‐expressing (LPV‐treated) cells with or without APH (0.4 µ m , 24 h) treatment. Scale bars: 10 µm. The number of 53BP1 bodies per nucleus was calculated. d) Immunofluorescence staining of lamin A/C in shCTRL and shZMPSTE24 cells. Scale bars: 10 µm. The percentage of micronuclei was calculated. n = 3. e) The proliferation of shCTRL and shZMPSTE24 cells was measured (left). The viability of shCTRL and shZMPSTE24 cells treated with the indicated concentrations of HU was measured (right). n = 3. f) Chromosomal aberrations, including breaks, gaps, rings, and dicentric chromosomes were analyzed for each metaphase in control and prelamin A‐expressing (LPV‐treated) cells with or without APH (0.3 µ m , 16 h) treatment. g) Chromosomal aberrations were analyzed for each metaphase in control and prelamin A‐expressing (LPV‐treated) cells with or without HU (4 mм, 4 h) and mirin (50 µ m , 4 h) treatment. Quantitative analysis results are shown as the mean ± SD. P values were determined by unpaired Student's t‐ test (a, b, d, and e) and the Mann‒Whitney test (c, f, and g). * p < 0.05; ** p < 0.01; *** p < 0.001.

Article Snippet: LPV (MCE, HY‐14588), FTI‐277 (MCE, HY‐15872A), HU (Sigma–Aldrich, H8627), APH (Glpbio, GC10867), colcemid (Glpbio, GC40664), Nutlin‐3A (MCE, HY‐10029), and mirin (MCE, HY‐117693) were used.

Techniques: Expressing, Staining, Knockdown, Western Blot, Immunofluorescence, Control

Journal: Advanced Science

Article Title: The Compromised Fanconi Anemia Pathway in Prelamin A‐Expressing Cells Contributes to Replication Stress‐Induced Genomic Instability

doi: 10.1002/advs.202307751

Figure Lengend Snippet: Downregulation of the FA/BRCA gene network in prelamin A‐expressing cells. a) The expression of FANCD2 and RAD51 in ZMPSTE24 ‐knockdown cells was measured by western blotting. n = 3. b) mRNA levels of FA/BRCA genes in ZMPSTE24 ‐knockdown cells were measured by qPCR. n = 3. c) Western blotting was used to measure the expression of FANCD2 and RAD51 in cells treated with FTI (3 µ m , 48 h), LPV (20 µ m , 6 days) or LPV + FTI. The farnesylated prelamin A generated by LPV treatment (lane 3) migrated more rapidly than the nonfarnesylated prelamin A generated by FTI treatment (lane 2 and lane 4). n = 3. d) The expression of RAD51 in asynchronous, S‐phase‐synchronized and M‐phase‐synchronized control and ZMPSTE24 ‐knockdown cells was measured by western blotting. Asyn, asynchronous. n = 3. e) The expression of FANCD2 in asynchronous, G1‐phase‐synchronized, S‐phase‐synchronized and M‐phase‐synchronized control and ZMPSTE24 ‐knockdown cells was measured by western blotting. Asyn, asynchronous. n = 3. f) SA‐β‐gal staining of IMR90 fibroblasts at early (P6) and late (P20) passages. Scale bar: 100 µm. g) The protein levels of prelamin A, FANCD2, FANCI and RAD51 in IMR90 fibroblasts at early (P6) and late (P20) passages were detected by western blotting. Quantitative analysis results are shown as the mean ± SD. P values were determined by unpaired Student's t‐ test (a, b, d, and e) and one‐way ANOVA (c). ns, not significant; * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001.

Article Snippet: LPV (MCE, HY‐14588), FTI‐277 (MCE, HY‐15872A), HU (Sigma–Aldrich, H8627), APH (Glpbio, GC10867), colcemid (Glpbio, GC40664), Nutlin‐3A (MCE, HY‐10029), and mirin (MCE, HY‐117693) were used.

Techniques: Expressing, Knockdown, Western Blot, Generated, Control, Staining