Journal: Frontiers in Cell and Developmental Biology

Article Title: Follicular fluid-derived exosomal LINC02701 promotes granulosa cell apoptosis through the GRP75–P53 axis in active endometriosis

doi: 10.3389/fcell.2026.1805254

Figure Lengend Snippet: LINC02701 weakens the GRP75–P53 interaction and enhances P53-associated apoptotic signaling. (A) Co-immunoprecipitation (Co-IP) assay validating the interaction between GRP75 and P53 in SVOG cells, with Input, IgG, and GRP75 IP controls shown. (B) Co-IP analysis of the GRP75–P53 interaction in LINC02701-overexpressing (OE) and untreated (WT) SVOG cells, with quantitative analysis of relative P53 expression in the immunoprecipitated fractions. (C) Immunofluorescence staining of P53 in Vector, WT, and OE cells, together with quantitative analysis of relative fluorescence intensity. Relative P53 fluorescence intensity was quantified using ImageJ after background subtraction from four random fields per condition in three independent experiments. Scale bar = 100 μm. (D) qPCR analysis of apoptosis-related genes, including BAX , PUMA , P21 , and BCL2 , in OE and WT cells. (E) Western blot analysis of BAX, PUMA, and P21 protein expression in OE and WT cells, together with densitometric quantification normalized to GAPDH. OE, LINC02701-overexpressing cells; WT, untreated cells; Vector, empty vector control cells; IP-OE, immunoprecipitated fraction from OE cells; IP-WT, immunoprecipitated fraction from WT cells. Error bars represent mean ± SD. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001; ns, not significant.

Article Snippet: To determine whether the pro-apoptotic effect of LINC02701 is mediated through P53 signaling, SVOG cells overexpressing LINC02701 were treated with the P53 inhibitor Pifithrin-α hydrobromide (MedChemExpress, China).

Techniques: Co-Immunoprecipitation Assay, Expressing, Immunoprecipitation, Immunofluorescence, Staining, Plasmid Preparation, Fluorescence, Western Blot, Control

Journal: Frontiers in Cell and Developmental Biology

Article Title: Follicular fluid-derived exosomal LINC02701 promotes granulosa cell apoptosis through the GRP75–P53 axis in active endometriosis

doi: 10.3389/fcell.2026.1805254

Figure Lengend Snippet: Inhibition of P53 attenuates LINC02701-induced apoptosis. (A) Representative flow cytometry plots of apoptosis in WT + vehicle, OE + vehicle, and OE + Pifithrin-α groups. (B) Quantification of apoptotic cells. (C) Western blot analysis of apoptosis-related proteins, including BAX, PUMA, and cleaved caspase-3. (D) Densitometric quantification of Western blot results. WT, untreated cells; OE, LINC02701-overexpressing cells. Error bars represent mean ± SD.

Article Snippet: To determine whether the pro-apoptotic effect of LINC02701 is mediated through P53 signaling, SVOG cells overexpressing LINC02701 were treated with the P53 inhibitor Pifithrin-α hydrobromide (MedChemExpress, China).

Techniques: Inhibition, Flow Cytometry, Western Blot

Journal: Frontiers in Cell and Developmental Biology

Article Title: Follicular fluid-derived exosomal LINC02701 promotes granulosa cell apoptosis through the GRP75–P53 axis in active endometriosis

doi: 10.3389/fcell.2026.1805254

Figure Lengend Snippet: Proposed mechanism by which follicular fluid-derived exosomal LINC02701 promotes granulosa-cell apoptosis in active endometriosis. Exosomal LINC02701 enters granulosa cells, interacts with GRP75, weakens the GRP75–P53 interaction, promotes P53 accumulation, and activates downstream apoptotic signaling.

Article Snippet: To determine whether the pro-apoptotic effect of LINC02701 is mediated through P53 signaling, SVOG cells overexpressing LINC02701 were treated with the P53 inhibitor Pifithrin-α hydrobromide (MedChemExpress, China).

Techniques: Derivative Assay

Journal: Genes

Article Title: CALB2 Expression Is Associated with Tumor Progression and Prognosis in Colorectal Adenocarcinoma

doi: 10.3390/genes17050510

Figure Lengend Snippet: Association of CALB2 expression with p53 signaling in colorectal adenocarcinoma. ( A , B ) Western blot analysis of p-p53-Ser33 and total p53 protein levels ( A ) and corresponding densitometric quantification ( B ) in HCT116 cells following CALB2 knockdown using two independent siRNAs (si1-CALB2 and si2-CALB2) or negative control (NC). ( C , D ) Western blot analysis of p-p53-Ser33 and total p53 protein levels ( C ) and corresponding densitometric quantification ( D ) in HT29 cells after CALB2 knockdown or negative control (NC). ( E , F ) Western blot analysis of p-p53-Ser33 and total p53 protein levels ( E ) and corresponding densitometric quantification ( F ) in SW48 cells after CALB2 overexpression (OE-CALB2) or negative control (NC). ( G , H ) Western blot analysis of p-p53-Ser33 and total p53 protein levels ( G ) and corresponding densitometric quantification ( H ) in SW480 cells after CALB2 overexpression (OE-CALB2) or negative control (NC). ( I – K ) RT-qPCR analysis of CALB2 ( I ), p21(CDKN1A) ( J ), and BAX ( K ) mRNA levels in HCT116 cells transfected with NC, si1-CALB2, or si1-CALB2 combined with the p53 inhibitor PFT-α (10 μM). ( L ) CCK-8 assay of HCT116 cells in the NC, si1-CALB2, and si1-CALB2+PFT-α groups over a 96 h period. ( M , N ) Representative images of scratch wound-healing assays ( M ) and quantitative analysis of relative migration rates ( N ) in HCT116 cells at 0, 24, and 48 h post-scratching. Scale bar = 250 μm. ( O – Q ) Representative double immunofluorescence staining ( O ) and quantitative analysis of fluorescence intensity ( P , Q ) for CALB2 and p-p53-Ser33 in xenograft tumor tissues from NC and OE-CALB2 groups. Scale bar = 50 μm. (n = 3, * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001).

Article Snippet: The p53-specific inhibitor PFT-α hydrobromide (PFT-α, 10 μM; MCE, Monmouth Junction, NJ, USA) was introduced to the culture medium 24 h after siRNA transfection.

Techniques: Expressing, Western Blot, Knockdown, Negative Control, Over Expression, Quantitative RT-PCR, Transfection, CCK-8 Assay, Migration, Double Immunofluorescence Staining, Fluorescence

Journal: bioRxiv

Article Title: Tau-induced mitochondrial reverse electron transport drives neurodegeneration

doi: 10.64898/2026.04.04.716514

Figure Lengend Snippet: a, Representative WB and quantification showing effect of VDAC, Hsp70, and Hsp90 inhibitor treatment on tau entry into mitochondria in the in vitro import assay. Mitochondrial p-tau level is normalized with the C-II protein SDHA. b , RET-ROS measurements after p-tau import into control or inhibitor-treated mitochondria. c , Representative WB and quantification showing total PHF-1 tau level in the in vitro import assay mixture. Total p-tau level is normalized with actin, which is known to be associated with mitochondria. d , RET-ROS measurements in control or inhibitor-treated mitochondria without p-tau import. e , WBs and quantification showing knockdown efficiency by VDAC1, Hsp70, and Hsp90 siRNAs. f , Representative WBs and quantification showing the effect of VDAC1, Hsp70, and Hsp90 siRNAs on tau entry into mitochondria in the in vitro import assay and total PHF-1 tau level in the in vitro import assay. g , h , RET-ROS measurements in mitochondria from control or siRNA treated cells with ( g ) or without ( h ) p-tau import into mitochondria. All data are means ± SEM; statistical significance was determined by one-way ANOVA with Dunnett’s multiple test ( a, b, c, d, f, g, h ), or two-tailed unpaired Student’s t test ( e ). *p<0.05; **p<0.01; ***p<0.001; ****p<0.0001. Each data point in a - h represents an independent experimental repeat.

Article Snippet: HEK293T cells were treated with the VDAC inhibitor DIDS (50 μM, Sigma, Cat#309795), Hsp70 inhibitor pifithrin-α hydrobromide (20μM, MedChemExpress, Cat#HY-15484), Hsp90 inhibitor Geldanamycin (1 μM, SelleckChem, Cat#S2713) for 6h.

Techniques: In Vitro, Control, Knockdown, Two Tailed Test

Journal: bioRxiv

Article Title: Tau-induced mitochondrial reverse electron transport drives neurodegeneration

doi: 10.64898/2026.04.04.716514

Figure Lengend Snippet: a, Representative WBs and quantification showing effect of VDAC, Hsp70, and Hsp90 inhibitors on the mitochondrial level of PHF-1 tau and total PHF-1 tau in the elav-GS>tau-R406W flies. In this GeneSwitch inducible model, tau-R406W expression is tightly controlled by the addition of RU486 to the fly food. Mitochondrial fractions or total cell lysates were used for WB, and mitochondrial or total PHF-1 p-tau was normalized by SDHA or actin. b , RET-ROS measurements in the mitochondria from control or inhibitor treated elav-GS>tau-R406W flies after tau induction by RU486. c - e , aversive taste memory assays in VDACi ( c ), Hsp70i ( d ), and Hsp90i ( e ) treated elav-GS>tau-R406W flies after tau induction by RU486. f , Climbing activity assay in VDACi, Hsp70i, and Hsp90i treated elav-GS>tau-R406W flies after tau induction by RU486. g , h , RET ( g ) and aversive taste memory ( h ) assays of control flies without tau transgene expression. All data are means ± SEM; statistical significance was determined by one-way ANOVA with Dunnett’s multiple test ( a, b, f, g ), or group analysis using multiple t test with Sidak-Bonferroni multiple comparison ( c , d , e , h ). *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001. Each data point represents independent experimental repeat. Three sets of flies with 10-12 flies in each set were used for the behavioral assays.

Article Snippet: HEK293T cells were treated with the VDAC inhibitor DIDS (50 μM, Sigma, Cat#309795), Hsp70 inhibitor pifithrin-α hydrobromide (20μM, MedChemExpress, Cat#HY-15484), Hsp90 inhibitor Geldanamycin (1 μM, SelleckChem, Cat#S2713) for 6h.

Techniques: Expressing, Control, Activity Assay, Comparison

Journal: bioRxiv

Article Title: Tau-induced mitochondrial reverse electron transport drives neurodegeneration

doi: 10.64898/2026.04.04.716514

Figure Lengend Snippet: a , Effect of H 2 O 2 and FK866 on the viability of tau-P301L hiPSC neurons and isogenic wildtype controls after 24h treatment, and the rescuing effect of CPT. b , Effect of H 2 O 2 and FK866 co-treatment on the viability of control and tau KD hiPSC neurons and the rescuing effect of CPT. c, d , Effect of DES treatment on the viability of control and tau KD hiPSC neurons ( c ) and the rescuing effect of CPT in DES (20 mM) treated control hiPSC neurons ( d ). e - g , WB and quantification showing the effect of inhibiting VDAC, Hsp70, or Hsp90 on PHF-1 tau entry into mitochondria without affecting total PHF-1 tau levels ( e ), and quantification of the effect of inhibitor treatment on RET activity ( f ) and stress sensitivity ( g ) of APP hiPSC neurons. h , Measurement of RET ROS and NAD + /NADH in purified mitochondria from control and tau-WT-EGFP or tau-S2A-EGFP-transfected normal hiPSC neurons. i , Representative images and quantification of p-S262 tau in tau-WT-EGFP or tau-S2A-EGFP transfected control hiPSC neurons with or without DES treatment. j , Measurement of RET ROS and NAD + /NADH in purified mitochondria from EGFP or MKI-EGFP transduced APP hiPSC neurons. k , Representative images and quantification of p-S262 tau in EGFP or MKI-EGFP transfected APP hiPSC neurons. l, m , Immunoblots ( l ) and quantification ( m ) showing effects of the various treatments on normalized levels of p-tau species in APP hiPSC neurons. n, o , Immunoblots ( n ) and quantification ( o ) showing effect of DES or DES/CPT co-treatment on normalized levels of p-tau species in control hiPSC neurons. All data are means ± SEM; statistical significance was determined by one-way ANOVA with Tukey’s post hoc test ( a, b, d, g, h, i, o ), two-way ANOVA with Tukey’s post hoc test ( c ), two-tailed unpaired Student’s t test ( j, k ), or one-way ANOVA with Dunnett’s multiple test ( e, f , m ). *p<0.05; **p<0.01; ***p<0.001; ****p<0.0001 ( a, b, c, d, g : n=6/group from 3 biological replicates and 2 wells/experiment; e, f, h, j, m, o : n=3 biological replicates; i, k : n=3 biological replicates, and each data point represents an average of 6 cells/experiment).

Article Snippet: HEK293T cells were treated with the VDAC inhibitor DIDS (50 μM, Sigma, Cat#309795), Hsp70 inhibitor pifithrin-α hydrobromide (20μM, MedChemExpress, Cat#HY-15484), Hsp90 inhibitor Geldanamycin (1 μM, SelleckChem, Cat#S2713) for 6h.

Techniques: Control, Activity Assay, Purification, Transfection, Western Blot, Two Tailed Test

Journal: bioRxiv

Article Title: Tau-induced mitochondrial reverse electron transport drives neurodegeneration

doi: 10.64898/2026.04.04.716514

Figure Lengend Snippet: a, Representative WBs and quantification showing effect of VDAC1, Hsp70, and Hsp90 lenti-shRNAs on the expression of the target proteins. b , Representative WBs and quantification showing effect of VDAC1, Hsp70, and Hsp90 lenti-shRNAs on the levels of mitochondrially localized PHF-1 tau and total PHF-1 tau in APP hiPSC neurons. Mitochondrial fractions or total cell lysates were used for WB, and mitochondrial or total PHF-1 p-tau was normalized by SDHA or actin. c , RET-ROS measurements in mitochondria from control or VDAC1, Hsp70, and Hsp90 lenti-shRNA treated APP hiPSC neurons. d , e , RET ( d ) and stress sensitivity ( e ) assays of control iPSC neurons treated with VDAC1, Hsp70, and Hsp90 inhibitors. All data are means ± SEM; statistical significance was determined by two-tailed unpaired Student’s t test ( a ) or one-way ANOVA with Dunnett’s multiple test ( b-e ). *p<0.05; **p<0.01; ***p<0.001; ****p<0.0001 ( e : n=6/group from 3 biological replicates and 2 wells/experiment).

Article Snippet: HEK293T cells were treated with the VDAC inhibitor DIDS (50 μM, Sigma, Cat#309795), Hsp70 inhibitor pifithrin-α hydrobromide (20μM, MedChemExpress, Cat#HY-15484), Hsp90 inhibitor Geldanamycin (1 μM, SelleckChem, Cat#S2713) for 6h.

Techniques: Expressing, Control, shRNA, Two Tailed Test