Journal: Membranes

Article Title: Role of Calcium-Activated Potassium Channels in Proliferation, Migration and Invasion of Human Chronic Myeloid Leukemia K562 Cells.

doi: 10.3390/membranes13060583

Figure Lengend Snippet: Figure 1. Ca2+-dependent potassium (KCa) channel expression and current-recording in K562 cells. (A) Representative whole-cell currents in the control conditions (pCa6) and after subsequent addition of apamin (300 nM, selective inhibitor of SK channels) and TRAM-34 (1 µM, inhibitor of IK channels), recorded in the range of holding membrane potentials from +20 mV to +80 mV, in 10 mV increments. (B) The mean (± SEM) whole-cell currents decreased after the subsequent addition of apamin and TRAM-34. Note that SE are within the size of the symbol. ****—significantly different for each condition at all membrane potentials, p < 0.0001. (C) RT-PCR analysis of KCa channel mRNA expression: M—size marker, KCNN2 (SK2)—423 bp, KCNN3 (SK3)—134 bp, KCNN4 (IK)—325 bp; KCNN1 (SK1, 282 bp) was not detected. Cropped gels are shown with enhanced contrast. Original gels are shown in Supplementary Figure S2. (D) Immunofluorescent staining confirmed the presence of SK2, SK3 and IK proteins in K562 cells. Blue channel—cell nuclei, red channel—SK2 and SK3, green channel—IK. No staining was observed when cells were incubated with only secondary fluorescent antibodies. Scale bar: 30 µM.

Article Snippet: For immunofluorescent staining, specific primary antibodies against the intracellular loop of SK2 (Alomone Labs, Jerusalem, Israel, cat. no. #APC-028), SK3 (Alomone Labs, Jerusalem, Israel, cat. no. #APC-025) channels and the extracellular loop of IK channels (conjugated to ATTO Fluor 488 dye, Alomone Labs, Jerusalem, Israel, cat. no. #ALM-051-AG) were used.

Techniques: Expressing, Control, Membrane, Reverse Transcription Polymerase Chain Reaction, Marker, Staining, Incubation

Journal: Membranes

Article Title: Role of Calcium-Activated Potassium Channels in Proliferation, Migration and Invasion of Human Chronic Myeloid Leukemia K562 Cells.

doi: 10.3390/membranes13060583

Figure Lengend Snippet: Figure 2. Pharmacological separation of SK2 and SK3 activity in the plasma membrane of K562 cells. (A) Representative whole-cell currents recorded in control conditions (pCa 6) and in the presence of 300 pM (blocks SK2 currents) or 8 nM (blocks SK3 currents) of apamin. (B) The mean (±SEM) whole-cell currents in control and after the addition of apamin (300 pM and 8 nM). Note that SE are within the size of the symbol. ****—significantly different for each condition at all membrane potentials, p < 0.0001.

Article Snippet: For immunofluorescent staining, specific primary antibodies against the intracellular loop of SK2 (Alomone Labs, Jerusalem, Israel, cat. no. #APC-028), SK3 (Alomone Labs, Jerusalem, Israel, cat. no. #APC-025) channels and the extracellular loop of IK channels (conjugated to ATTO Fluor 488 dye, Alomone Labs, Jerusalem, Israel, cat. no. #ALM-051-AG) were used.

Techniques: Activity Assay, Clinical Proteomics, Membrane, Control

Journal: Membranes

Article Title: Role of Calcium-Activated Potassium Channels in Proliferation, Migration and Invasion of Human Chronic Myeloid Leukemia K562 Cells.

doi: 10.3390/membranes13060583

Figure Lengend Snippet: Figure 6. The role of KCa channels in proliferation, migration and invasion of human CML K562 cells. Functional expression of SK2, SK3 and IK channels was shown using PCR, immunofluorescence and electrophysiological assay, using selective channel inhibitors (apamin and TRAM-34). Blocking of IK- and SK-mediated K+ currents resulted in a decrease in Ca2+ influx and reduced the proliferative, invasive and migratory activities of K562 cells.

Article Snippet: For immunofluorescent staining, specific primary antibodies against the intracellular loop of SK2 (Alomone Labs, Jerusalem, Israel, cat. no. #APC-028), SK3 (Alomone Labs, Jerusalem, Israel, cat. no. #APC-025) channels and the extracellular loop of IK channels (conjugated to ATTO Fluor 488 dye, Alomone Labs, Jerusalem, Israel, cat. no. #ALM-051-AG) were used.

Techniques: Migration, Functional Assay, Expressing, Blocking Assay

Journal: Molecular Cell

Article Title: BAP1 enhances Polycomb repression by counteracting widespread H2AK119ub1 deposition and chromatin condensation

doi: 10.1016/j.molcel.2021.06.020

Figure Lengend Snippet:

Article Snippet: Western blot and ChIP analyses were performed with: anti-BAP1 (D7W70; Cell Signaling), anti-Suz12 (D39F6; Cell Signaling Technology), anti-FLAG (F3165; Sigma-Aldrich), anti-H3K27me3 (9733; Cell Signaling Technology), anti-H2AK119ub1 (8240; Cell Signaling Technology), anti-H3 (ab1791; Abcam), anti-H2A (07-146; Sigma-Aldrich), anti-RING1B (homemade ( )), anti-PCGF1, anti-PCGF2, and anti-PCGF6 (Pasini’s lab, homemade ( )), anti-JARID2 (13594, Cell Signaling Technology), anti-AEBP2 (14129; Cell signaling Technology), anti-HA (C29F4, Cell Signaling Technology), anti-ASXL2 (A302-037A, Bethyl), anti-ASXL1 (D1B6V, Cell Signaling Technology) anti-KDM1B/LSD2 (E1R60, Cell Signaling Technology), anti-OGT (SC-32921, Santa-Cruz), anti-HCFC1 (A301-400A, Bethyl), anti-FOXK2 (A301-730A, Bethyl), anti-MBD6 (SAB1305225-40TST, Sigma-Aldrich), anti-H3K27ac (AB4729, Abcam) anti-PCGF3/5 (AB201510, Abcam), anti-UTX (Helin lab, homemade ( )), anti-MTF2 (Proteintech 16208-1-AP), anti-CBX7 (abcam, ab21873), anti-RYBP/DEDAF (Sigma-Aldrich, PRS2227), anti-H3K36me2 (abcam, ab9049), anti-RING1B-pS168 (abcam, ab234421).

Techniques: Control, Virus, Recombinant, Transfection, Purification, Extraction, Sequencing, Imaging, Plasmid Preparation, Software