Journal: Blood Cancer Journal

Article Title: Downregulation of PA28α induces proteasome remodeling and results in resistance to proteasome inhibitors in multiple myeloma

doi: 10.1038/s41408-020-00393-0

Figure Lengend Snippet: a Western blot analysis of ubiquitinated proteins in total protein lysates extracted from LP1 and RPMI8226 PA28α knockdown stable cells. β-actin as a loading control. b OPP pulse-chase assay of proteasome degradation and protein synthesis in LP1 PA28α knockdown stable cells. Western blot analysis of immunoglobulin lambda light chain (ƛ IgL) ( c ), eIF2α, p62/SQSTM1, and LC3B ( d ) in LP1 and RPMI8226 PA28α knockdown stable cells, β-actin as a loading control. ** P < 0.01, Student’s t test. e Working model of PA28α knockdown in MM.

Article Snippet: Antibodies used were as follows: PA28α (Cell Signaling), PSMA2 (Cell Signaling), S5a (Cell signaling), PA28β (Cell Signaling), Phospho-eIF2α (Ser51) (Cell signaling), eIF2α (Cell signaling), α-tubulin (Genetex), PA28γ (Genetex), PSMB5 (Genetex), PSMB6 (Enzo life science), PSMB7 (Genetex), PSMB8 (Genetex), PSMB9 (R&D systems), PSMB10 (R&D systems), Rpt5 (Enzo life science), Ubiquitin (Cell Signaling), β-actin (Santa Cruz Technology), TCF11/NRF1 (Cell Signaling), LC3B (Cell Signaling), p62/SQSTM1 (MBL International) human Ig lambda light chain (R&D systems), actin (Sigma). pLKO.1 empty vector, shRNA vector targeting human PA28α, NRF1 siRNA (ON-TARGETplus SMARTpool), PA28α siRNA (Accell SMARTpool), and control siRNA were purchased from Dharmacon.

Techniques: Western Blot, Knockdown, Control, Pulse Chase

Journal: Cell

Article Title: B cell genomics behind cross-neutralization of SARS-CoV-2 variants and SARS-CoV

doi: 10.1016/j.cell.2021.04.032

Figure Lengend Snippet:

Article Snippet: HRP Conjugated Goat anti-Human Lambda Light Chain Antibody , Bio-Rad , Cat#STAR129P; RRID: AB_1102721.

Techniques: Virus, Recombinant, Staining, Plasmid Preparation, Expressing, Modification, Luciferase, Cell Culture, Lysis, Reporter Gene Assay, Enzyme-linked Immunosorbent Assay, Electron Microscopy, Cloning, Software, Chromatography, Spectrophotometry

Journal: Blood

Article Title: Myeloma cell–derived Runx2 promotes myeloma progression in bone

doi: 10.1182/blood-2014-12-613968

Figure Lengend Snippet: Runx2 knockdown inhibits MM progression in vivo. (A) Expression of Runx2 in 2 Runx2 k/d 5TGM1 cell lines (Runx2 shRNA #90– and Runx2 shRNA #91–transfected 5TGM1 cells) was probed by western blot. Both Runx2 shRNA #90 and #91 decreased the expression of Runx2 compared with NT control. (B) Serum IgG2bκ was measured by ELISA 6 weeks after IV injection of NT control or Runx2 k/d 5TGM1 cells. Error bars represent mean ± SEM (n = 10 animals per group). Significant differences between groups are indicated by P value. (C) H&E-stained bone sections from mice injected IV with either NT control or Runx2 k/d 5TGM1 cells. Tumors were present in mice injected with NT cells, but not in the mice injected with Runx2 k/d cells (original magnification, ×100). Inset, Abundant myeloma cells in NT-bearing mice compared with Runx2 k/d 5TGM1 cell-injected mice. (D) Survival was significantly increased in mice injected IV with 5TGM1 Runx2 k/d cells (clone #90 and #91) compared with those injected with NT 5TGM1 cells. (E) Six weeks after intratibial injection of Runx2 k/d or NT 5TGM1 cells, levels of serum IgG2bκ were measured by ELISA. Error bars represent mean ± SEM (n = 10 animals per group). Significant differences between groups are indicated by P value. (F) Western blot shows reduction of Runx2 expression in MM.1R cells transduced with Runx2 shRNA compared with wild-type (WT) or NT control cells. (G) Six weeks after s.c. injection of NT or Runx2 k/d human MM.1R cells, serum human Ig λ light chain was measured by ELISA. Error bars represent mean ± SEM (n = 10 animals per group). Significant differences between groups are indicated by P value.

Article Snippet: The levels of human Ig λ light chain or mouse IgG2bκ in mouse sera were measured using human Ig λ or mouse IgG2bκ ELISA kits (Bethyl Laboratories Inc), respectively.

Techniques: Knockdown, In Vivo, Expressing, shRNA, Transfection, Western Blot, Control, Enzyme-linked Immunosorbent Assay, IV Injection, Staining, Injection, Transduction

Journal: Cells

Article Title: Empagliflozin Preserves Cardiomyocyte Structural Homeostasis via the Stabilization of the Integrin α5–Desmocollin-2 Adhesion Axis in Sepsis-Induced Cardiomyopathy

doi: 10.3390/cells14181452

Figure Lengend Snippet: Empagliflozin ameliorates LPS-induced cardiac dysfunction and myocardial injury. ( A ) Echocardiography panels: top, B-mode long-axis image; middle, M-mode from the parasternal short-axis at the papillary muscle level; bottom, pulsed-wave Doppler of transmitral inflow (E and A waves). ( B ) Quantification of cardiac function parameters: Ejection Fraction (EF), Fractional Shortening (FS), and E/A Ratio. ( C ) Representative images of Hematoxylin and Eosin (H & E) staining of myocardial tissue sections showing inflammatory infiltration. Longitudinal and cross sections are shown with a scale bar of 1 mm. Magnified views of perivascular and interstitial regions are shown with a scale bar of 20 µm; green arrows indicate inflammatory cells. ( D ) Representative images of Masson’s trichrome staining revealing myocardial fibrosis. Longitudinal sections are shown with a scale bar of 1 mm, and cross sections with a scale bar of 0.5 mm. Magnified views of perivascular and interstitial regions are shown with a scale bar of 40 µm; yellow arrows indicate collagen deposition (fibrotic areas). ( E ) Quantification of the fibrotic area from Masson’s trichrome staining. ( F ) Serum concentrations of cardiac troponin T (cTnT) and C-reactive protein (CRP) measured by ELISA. Data are presented as mean ± SEM. Statistical analysis was performed using one-way ANOVA. **** p < 0.0001, *** p < 0.001, ** p < 0.01.

Article Snippet: Serum levels of cardiac troponin T (cTnT) and C-reactive protein (CRP) were quantified using commercial enzyme-linked immunosorbent assay (ELISA) kits (cTnT: Cat. No. orb565454, Biorbyt, Wuhan, China; CRP: Cat. No. orb219654, Biorbyt, Wuhan, China) according to the manufacturer’s instructions.

Techniques: Staining, Enzyme-linked Immunosorbent Assay