Journal: bioRxiv

Article Title: Complement inhibition by C3-siRNA treatment prevents AChR loss and reduces complement activation in the rat Passive Transfer Myasthenia Gravis (PTMG)

doi: 10.1101/2025.08.31.673367

Figure Lengend Snippet: A) After baseline measurements on day 0, subcutaneous treatment injections were administered on day 1 and 8. Intermediate testing and blood collection were performed on day 4, 7 and 11 (not shown). On day 15, indicated with a red arrow, PTMG was induced using mAb35, while non-MG animals received a saline injection of the same volume. B-C) Plasma C3 expression was reduced by the sc administration of C3-siRNA at various dose levels in B) non-MG animals and C) PTMG animals. For animals receiving 2 x 30 mg/kg treatment, doses were administered on days 1 and 8. For animals receiving 1 x 10 or 1 x 30 mg/kg doses, administration occurred on day 8, as indicated by the black arrows on the graphs. Saline-treated animals showed no significant change in C3 levels over time. Group averages are shown. D) Plasma C3 level changes in individual treatment groups on day 17, normalized to baseline levels on day 0. Individual plasma C3 levels per animal are shown. Statistical analysis was performed with Two-Way ANOVA (B-D), followed by One-Way ANOVA and post-hoc Bonferroni’s multiple comparison test (D); α=0.05; ****p<0.0001. A) Created in BioRender. Schöttler, A. (2025) https://BioRender.com/b4xxodd .

Article Snippet: PTMG was induced at 11-weeks of age by sc injection of 40 pmol/100 g body weight of mAb35 [deposited to the DSHB by Lindstrom, J. (Developmental Studies Hybridoma Bank (DSHB) product mAb35) on day 15, after C3-siRNA treatment.

Techniques: Saline, Injection, Clinical Proteomics, Expressing, Comparison

Journal: bioRxiv

Article Title: Complement inhibition by C3-siRNA treatment prevents AChR loss and reduces complement activation in the rat Passive Transfer Myasthenia Gravis (PTMG)

doi: 10.1101/2025.08.31.673367

Figure Lengend Snippet: A) After baseline measurements on day 0, subcutaneous treatment injections were administered on day 1 and 8. Intermediate testing and blood collection were performed on day 4, 7 and 11 (not shown). On day 15, indicated with a red arrow, PTMG was induced using mAb35, while non-MG animals received a saline injection of the same volume. B-C) Plasma C3 expression was reduced by the sc administration of C3-siRNA at various dose levels in B) non-MG animals and C) PTMG animals. For animals receiving 2 x 30 mg/kg treatment, doses were administered on days 1 and 8. For animals receiving 1 x 10 or 1 x 30 mg/kg doses, administration occurred on day 8, as indicated by the black arrows on the graphs. Saline-treated animals showed no significant change in C3 levels over time. Group averages are shown. D) Plasma C3 level changes in individual treatment groups on day 17, normalized to baseline levels on day 0. Individual plasma C3 levels per animal are shown. Statistical analysis was performed with Two-Way ANOVA (B-D), followed by One-Way ANOVA and post-hoc Bonferroni’s multiple comparison test (D); α=0.05; ****p<0.0001. A) Created in BioRender. Schöttler, A. (2025) https://BioRender.com/b4xxodd .

Article Snippet: PTMG was induced at 11-weeks of age by sc injection of 40 pmol/100 g body weight of mAb35 [deposited to the DSHB by Lindstrom, J. (Developmental Studies Hybridoma Bank (DSHB) product mAb35) on day 15, after C3-siRNA treatment.

Techniques: Saline, Injection, Clinical Proteomics, Expressing, Comparison

Journal: The Journal of Biological Chemistry

Article Title: Novel markers of MCL1 inhibitor sensitivity in triple-negative breast cancer cells

doi: 10.1016/j.jbc.2024.107375

Figure Lengend Snippet: A 4 gene signature (GS) correlates with AZD5991 sensitivity and MCL-1 dependency in TNBC cell lines. A , analysis of gene expression correlation with TIMER software in 139 TNBC tumors (TCGA) showed that the 4 genes positively correlate with each other. B , 18 TNBC cell lines from GDSC are divided into AZD5991 resistant ( green ) and sensitive ( yellow ) groups. mRNA levels (RPKM) of AXL, ETS1, IL6, and EFEMP1 in all cell lines were extracted from CCLE RNAseq database. The median level of each gene is calculated and listed on top. Gene levels above the median is marked in red and scored 1 and below median is scored 0. The sum of the 4 gene scores is listed on right . C , correlation of GS scores with AZD5991 sensitivity was analyzed with ROC curve with sensitive cells defined as event 1. The results showed a significant correlation ( p = 0.002) with an ROC area 0.95 (ROC area 1.0 indicates perfect correlation). D, 13 TNBC cell lines with MCL-1 CRISPR (DepMap public 23Q2) are divided into MCL-1 dependent ( green ) and independent ( yellow ) groups based on their Chromos scores. Gene scores are calculated as described in B. Correlation of GS scores with MCL-1 dependency was analyzed with ROC curve ( E ). The results showed a significant correlation ( p = 0.04) with a ROC area 0.79).

Article Snippet: Antibodies to BCL2 (124), MCL1 (D35A5), pAKT (S473) (D9E), AKT (11E1), pJAK3 (Y980/981) (D44E3), JAK3 (D7B12), pSTAT3 (Y705) (D3A7), STAT3, pERK1/2 (T202/Y204) (197G2), ERK1/2 (137F5), AXL (C89E7) and ETS1 (D808A) were from Cell Signaling; EFEMP1 (fibulin-3, mAb35) and β-actin (C4) antibodies were from Santa Cruz.

Techniques: Expressing, Software, CRISPR

Journal: The Journal of Biological Chemistry

Article Title: Novel markers of MCL1 inhibitor sensitivity in triple-negative breast cancer cells

doi: 10.1016/j.jbc.2024.107375

Figure Lengend Snippet: Inhibition of ETS1 reduces AXL, IL6, and EFEMP1 expression and increases MCL1 inhibitor sensitivity via expression of BCL-2. A , HS578T and MDA231 cells were transfected with control siRNA or ETS1 siRNA for 24 h. B, HS578T and MDA231 cells were treated with vehicle or TK216 (1 μM) for 24 h. mRNA were qPCR analyzed for the indicated genes. Average (triplicate) relative mRNA is presented with SD indicated. Asterisks indicate statistical significance. Representative results of two independent experiments. C , HS578T and MDA231 cells were treated with vehicle, TK216 (1μM), S63845 (1μM), or TK216 + S63845 for 3 days and then analyzed for sub-G1 cells. % sub-G1 cells are presented. Asterisks indicate statistical significance. Representative results of three independent experiments. D , MDA231 cells were treated with vehicle, S63845 (1μM), or S63845 plus BGB324 (1 μM) or TK216 (1μM), for 24 h. Lysates were immunoblotted for the indicated proteins. Representative results of two independent experiments. Original images are presented in Fig. S5 . E , The cells were treated with vehicle or the ERK inhibitor ulixertinib (2 μM) for 24 h. Lysates were immunoblotted for the indicated proteins. Representative results of two independent experiments. Original images are presented in Fig. S6 . F , MDA231 cells were treated with vehicle, S63845 (1 μM), or S63845 plus ABT-199 (2 μM and 5 μM) for 72 h. G , The cells were transfected with control siRNA or the indicated siRNAs against AXL, ETS1, IL6, or EFEMP1 and then treated with vehicle or S63845 for 72 h. Cells were analyzed with FACS for the cell cycle. The average % Sub-G1 cells were presented with SD indicated. Asterisks indicate statistical significance. Representative results of two independent experiments.

Article Snippet: Antibodies to BCL2 (124), MCL1 (D35A5), pAKT (S473) (D9E), AKT (11E1), pJAK3 (Y980/981) (D44E3), JAK3 (D7B12), pSTAT3 (Y705) (D3A7), STAT3, pERK1/2 (T202/Y204) (197G2), ERK1/2 (137F5), AXL (C89E7) and ETS1 (D808A) were from Cell Signaling; EFEMP1 (fibulin-3, mAb35) and β-actin (C4) antibodies were from Santa Cruz.

Techniques: Inhibition, Expressing, Transfection, Control

Journal: The Journal of Biological Chemistry

Article Title: Novel markers of MCL1 inhibitor sensitivity in triple-negative breast cancer cells

doi: 10.1016/j.jbc.2024.107375

Figure Lengend Snippet: Overexpression of ETS1 and exogenous EFEMP1 and IL6 increases BCL2 expression and promotes MCL1 inhibitor resistance. A , control BT20 cells and BT20-pLIX-ETS1 cells were treated with vehicle or doxycycline (DOX, 0.5 μg/ml) for 24 h. Lysates were immunoblotted for the indicated proteins. Representative images of two independent experiments. B , control BT20 cells and BT20-pLIX-ETS1 cells were treated with vehicle or doxycycline (DOX, 0.5 μg/ml) in combination with vehicle or S63845 for 72 h. Cells were analyzed with FACS for cell cycle. Average % Sub-G1 cells (apoptosis) were presented with SD indicated. Asterisks indicate statistical significance. Representative results of two independent experiments. BT20 cells were FBS-starved for 24 h and then treated with vehicle or recombinant EFEMP1 (50 ng/ml) for 30 min ( C ) or recombinant IL6 (50 ng/ml) for 10 min ( D ). E , BT20 cells were treated with vehicle or S63845 (1 μM) or S63845 in combination with EFEMP1 plus IL6 for 24 h. Lysates were immunoblotted for the indicated proteins. Representative results of two independent experiments. F , BT20 cells were treated with vehicle or S63845 (1 μM) or S63845 in combination with EFEMP1 plus IL6 for 72 h. Cells were analyzed with FACS for the cell cycle. Average % Sub-G1 cells (apoptosis) were presented with SD indicated. Asterisks indicate statistical significance. Representative results of two independent experiments.

Article Snippet: Antibodies to BCL2 (124), MCL1 (D35A5), pAKT (S473) (D9E), AKT (11E1), pJAK3 (Y980/981) (D44E3), JAK3 (D7B12), pSTAT3 (Y705) (D3A7), STAT3, pERK1/2 (T202/Y204) (197G2), ERK1/2 (137F5), AXL (C89E7) and ETS1 (D808A) were from Cell Signaling; EFEMP1 (fibulin-3, mAb35) and β-actin (C4) antibodies were from Santa Cruz.

Techniques: Over Expression, Expressing, Control, Recombinant